Research ArticleThe Combined Extract of Purple Waxy Cornand Ginger Prevents Cataractogenesis and Retinopathy inStreptozotocin-Diabetic Rats

Paphaphat Thiraphatthanavong12 Jintanaporn Wattanathorn23

Supaporn Muchimapura23 Wipawee Thukham-mee23

Kamol Lertrat4 and Bhalang Suriharn4

1Department of Physiology and Graduate School (Neuroscience Program) Faculty of MedicineKhon Kaen University Khon Kaen 40002 Thailand2Integrative Complementary Alternative Medicine Research and Development Center Khon Kaen UniversityKhon Kaen 40002 Thailand3Department of Physiology Faculty of Medicine Khon Kaen Unieversity Khon Kaen 40002 Thailand4Faculty of Agriculture Khon Kaen University Khon Kaen 40002 Thailand

Correspondence should be addressed to Jintanaporn Wattanathorn jintanapornwyahoocom

Received 20 October 2014 Revised 12 December 2014 Accepted 14 December 2014 Published 31 December 2014

Academic Editor Jing Yi

Copyright copy 2014 Paphaphat Thiraphatthanavong et al This is an open access article distributed under the Creative CommonsAttribution License which permits unrestricted use distribution and reproduction in any medium provided the original work isproperly cited

Based on the crucial roles of oxidative stress and aldose reductase on diabetic complications and the protective effect against diabeticeye complication of purple waxy corn and ginger (PWCG) together with the synergistic effect concept we aimed to determineanticataract and antiretinopathy effects of the combined extract of purple waxy corn and ginger (PWCG) The streptozotocindiabetics with the blood glucose levels gt250mgsdotdLminus1 were orally given the extract at doses of 50 100 and 200mgkgsdotBWminus1 for10 weeks Then lens opacity and histopathology of retina were determined The changes of MDA together with the activities ofSOD CAT GPx and AR in lens were also determined using biochemical assays All doses of PWCG decreased lens opacity MDAand AR in the lens of diabetic rats The elevation of CAT and GPx activities was also observed The antiretinopathy property of thecombined extract was also confirmed by the increased number of neurons in ganglion cell layer and thickness of total retina andretinal nuclear layer in diabetic rats PWCG is the potential functional food to protect against diabetic cataract and retinopathyHowever further studies concerning toxicity and clinical trial are still essential

1 Introduction

Diabetic cataract and diabetic retinopathy are chronic pro-gressive potentially sight-threatening diseases which areassociated with hyperglycemia [1 2] The epidemiologicalstudy has shown that the prevalences of both cataractand retinopathy in diabetic patients are increased [3 4]accompanied with the increased diabetic patients worldwideThe effective treatment of diabetic cataract and diabeticretinopathy requires the skillful physician Therefore theseconditions still induce the important health problems in

the developing countries especially in a rural area so theprevention strategy is very much important

Recent findings have demonstrated that the increasedaldose reductase activity and the elevated oxidative stresscontribute an important role on the development of cataractand retinopathy in diabetesmellitus [5ndash8] Anthocyanins richdiet such as purple waxy corn and colored rice could preventdiabetic cataract [9 10] It has been reported that purple waxycorn extract decreases oxidative stress in lens of diabetic ratsresulting in the decreased cataractogenesis [9] In additionginger extract also exerts anticataractogenesis [11 12] Our

Hindawi Publishing CorporationOxidative Medicine and Cellular LongevityVolume 2014 Article ID 789406 11 pageshttpdxdoiorg1011552014789406

2 Oxidative Medicine and Cellular Longevity

pilot data show that the antioxidant effect of the combinationof purple waxy corn and ginger (PWCG) is more potent thanthe antioxidant effect of either purple waxy corn or gingeralone (pilot data) However all of the evidence mentionedearlier is the in vitro studyTherefore we aimed to determinethe in vivo effect of PWCG on cataract retinopathy oxidativestress and aldose reductase in lens of diabetic rats

2 Materials and Methods

21 Plant Material and Extract Preparation The plant mate-rials used in this study were dried seeds of purple waxy cornor Zea mays L (purple color KKU open pollinated cultivar)and rhizomes of ginger or Zingiber officinale Roscoe Theywere harvested during September 2012 and authenticated byAssociate Kamol Lertrat and Dr Bhalang Suriharn Facultyof Agriculture Khon Kaen University Khon Kaen ThailandThe voucher specimens (voucher specimen 2012001 and2012002) were kept at the Integrative Complementary Alter-native Medicine Research and Development Center KhonKaen University The dried seeds of purple waxy corn andrhizomes of ginger were extracted with 50 hydroalcoholicsolvent by maceration method at a ratio of 2 5 and 1 5(weight volume) respectively The samples were maceratedat room temperature for 3 days Both of the yielded extractswere concentrated by lyophilization and kept at 4∘C forfurther study The percentage yields of purple waxy cornand ginger extracts were 572 and 2526 respectively Thecombination extract was prepared by mixing the purplewaxy corn and ginger extracts at a ratio of 1 4 (this ratioprovided the highest anticataract potential) The contentsof total phenolic compounds and anthocyaninsin the com-bination extract were 4482 plusmn 237mgL GAEmg extractand 39242plusmn003mgL cyanidin-3-glucoside equivalentsmgextract respectively

22 Experimental Design The experimental animals usedin this study were male Wistar rats at the weight of 200ndash250 g (119899 = 8 per group) The animals were maintainedand treated in accordance with the guideline and approvalof the Ethical Committee on Animals Experiments of KhonKaenUniversity (AEKKU982555) All rats were divided intovarious groups as follows

Group I Control group all rats in this group were adminis-tered citrate buffer a vehicle of streptozotocin (STZ)

Group II DM + vehicle group the animals in this groupwere induced diabetesmellitus via single injection of STZ andreceived vehicle of the extract or distilled water

Group IIIndashV The animals in this group had DM + thecombination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW respectively

All rats in groups IIndashV were induced diabetes cataractby a single injection of STZ which was dissolved in citratebuffer (pH 45) at dose of 55mgsdotkgminus1 BW The animalswhich showed the blood sugar levels more than 250mgsdotdLminus1were recruited for further study All rats were treated with

the assigned interventions once daily at 3 days after injectionof STZ and maintained for 10 weeks Lens opacity wasevaluated every week using slit lamp microscope At the endof study lens were collected and the level of malondialdehyde(MDA) and the activities of scavenging enzymes activitiesincluding superoxide dismutase (SOD) catalase (CAT) andglutathione peroxidase (GPx) in the lens together with lenshistomorphology were determined

23 Determination of Fasting Blood Glucose Level Afterfasting overnight blood was collected from rat tails andfasting blood glucose levels were determined using ACCU-CHEK active This process was performed at the first weekand every 5 weeks throughout the study period

24 Cataract Evaluation via Slit Lamp Rat eyes were eval-uated every week using slit lamp microscope (DIOPTRIX-HAWKEYE France) by trained observer who was blind tothe treatment The severity of cataract was graded as 5 stagesaccording to the method of Suryanarayana and coworkers[13] as follows

Stage 0 Lens were clear and no vacuoles were observed

Stage 1 Vacuoles cover approximately one-half of the surfacesof the anterior pole forming a subcapsular cataract

Stage 2 Some vacuoles have disappeared and the cortexexhibits a hazy opacity

Stage 3 A hazy cortex remains and dense nuclear opacity ispresent

Stage 4 A mature cataract is observed as a dense opacity inboth cortex and nucleus

Data were presented as opacity index which was calcu-lated from the following formula

Opacity index

=

(number of eyes in each stage times stage of the eye)total number of eyes within group

(1)

25 Homogenate Preparation At the end of experimentlens were collected and homogenized in 10wv of 01Mphosphate buffer pH 74 containing 1mM EDTA Then thehomogenate was centrifuged at 10000 g at 4∘C for 1 hour andthe supernatantwas separated and used for the determinationof biochemical parameters

26 Determination of Malondialdehyde (MDA) Level Levelof malondialdehyde (MDA) a lipid peroxidationmarker wasmonitored by using thiobarbituric acid reacting substances(TBARS) assay In brief 100 120583L of sample was mixed withthe solution containing 100120583L of 81 (wv) sodium dodecylsulphate 750120583L 20 (vv) acetic acid (pH 35) and 750 120583L of08 thiobarbituric acid (TBA) The solution was heated ina water bath at 95∘C for one hour and cooled immediately

Oxidative Medicine and Cellular Longevity 3

under running tap water Then 500120583L chilled water and2500 120583L of butanol and pyridine [15 1 vv] were added intoeach tube and mixed thoroughly with vortex Then thesolutionwas centrifuged at 800timesg for 20minTheupper layerwas separated and absorbance was measured at 532 nm 133-Tetra ethoxy propane (TEP) was used as the reference [14]The level of MDA was expressed as Umg protein

27 Superoxide Dismutase (SOD) Assay The determinationof SOD activity was carried out via nitro blue tetrazolium(NBT) reduction assay In this assay the xanthine-xanthineoxidase system was used as a superoxide generator In briefthe reaction mixture contained 20 120583L of sample and 200 120583Lof reaction mixture consisting of 57mM phosphate buffersolution (KH

2

PO4

) 01mM EDTA 10mM cytochrome Csolution and 50 120583M of xanthine solution and 20 120583L ofxanthine oxidase solution (090mUmL) were prepared at25∘C The optical density was measured at 415 nm A systemdevoid of enzyme served as the control and three parallelexperimentswere conducted [15] SODactivitywas expressedas Umg protein

28 Catalase (CAT) Assay Lens catalase activity was deter-mined based on the ability of the enzyme to break downH2

O2

In brief 10 120583L of sample was mixed with the reactionmixture containing 50 120583L of 30mM hydrogen peroxide (in50mMphosphate buffer pH 70) 25120583L of H

2

SO4

and 150 120583Lof KMnO

4

After mixing thoroughly the optical densitywas measured at 490 nm A system devoid of the substrate(hydrogen peroxide) served as the control The difference inabsorbance per unit time was expressed as the activity Theamount of enzyme which is required to decompose 10M ofhydrogen peroxide per minute at pH 70 and 25∘ is regardedas one unit [16] The value of CAT activity was expressed asUmg protein

29 Glutathione Peroxidase (GPx) Assay This assay was per-formed based on the glutathione recycling method by using551015840-dithiobis (2-nitrobenzoic acid) (DTNB) and glutathionereductase According to this method the reaction betweenDTNB and GSH gave rise to the generation of 2-nitro-5-thiobenzoic acid and GSSG Since 2-nitro-5-thiobenzoicacid was a yellow colored product GSH concentrationcould be determined by measuring absorbance at 412 nmIn brief a mixture containing a 20 120583L of sample and thereaction mixture consisting of 10 120583L of dithiothreitol (DTT)in 667mM potassium phosphate buffer (pH 7) 100120583L ofsodium azide in 667mM potassium phosphate buffer (pH7) 10 120583L of glutathione solution and 100 120583L of hydrogenperoxide was mixed thoroughly and incubated at roomtemperature for 5ndash10 minutes Then 10 120583L of DTNB (55-dithiobis-2-nitrobenzoic acid) was added and the opticaldensity at 412 nmwas recorded at 25∘Cover a period of 5minActivitieswere expressed as nmolesminmg lens protein [17]GPx activity was expressed as Umg protein

210 Aldose Reductase (AR) Activity Assay Aldose reductaseactivity was evaluated using spectrophotometric method

An assay mixture containing 07mL of phosphate buffer(0067mol) 01mL of NADPH (25 times 10minus5mol) 01mL of DL-glyceraldehyde (substrate 5 times 10minus4mol) and 01mL of lenssupernatant was prepared Absorbance was recorded againsta reference cuvette containing all other components exceptthe substrate DL-glyceraldehydeThe final pH of the reactionmixture was adjusted to pH = 62 The determination wasperformed after adding the substrate or DL-glyceraldehydeby measuring the decrease in NADPH absorbance at 390 nmover a 4-minute period [18] The enzyme activity wasexpressed as (nmolminmg)

211 Histopathological Analysis of Rat Lens and Retina Theeye balls from five rats per group were fixed in 10 formalinovernight embedded in paraffin sectioned at 5 120583m thick andstained with hematoxylin and eosin Light microscope wasused to evaluate the histomorphology of lens The severityof histomorphological change of lens was graded as a 5-grade score according to the method of Agarwal et al [19]as described as follow

Grade 0 Presence of anterior epithelium with lens fibers

Grade 1 Presence of anterior epithelium lens fibers andvacuoles

Grade 2 Presence of anterior epithelium lens fibers vacuolesand homogenized area

Grade 3 Absence of anterior epithelium presence of lensfibers vacuoles and homogenized area

Grade 4 Presence of lens fibers and homogenized area onlyHistomorphological changes of retina including the total

retinal thickness (from inner limiting membrane to Bruchrsquosmembrane) the thickness of the retinal outer nuclear layerand the number of cells in the ganglion cell layer were alsoperformed The average thickness of retina was evaluatedusing 3 adjacent fields and total five images in each group [20]The results were showed as mean plusmn SEM

212 Statistical Analysis All data were presented as mean plusmnstandard error mean (mean plusmn SEM) The analysis of datawas performed using one-way analysis of variance (ANOVA)followed by the post hoc test of LSD via SPSS version 15Statistical differences were considered at 119875 value lt 005

3 Results

31 Effect of PWCG on Fasting Blood Sugar Level The effectof PWCG on the average fasting blood glucose levels wasdetermined and the results were shown in Figure 1 Theresults showed that diabetic rats which received distilledwater or vehicle used in this study significantly enhanced theblood sugar levels throughout the study period (119875 value lt0001 all compared with control rats which received distilledwater) It was found that the blood sugar levels of all diabeticrats were more than 300mgsdotdLminus1 throughout the study

4 Oxidative Medicine and Cellular Longevity

Table 1 Opacity index of the lens of normal and diabetic rats (DM) which received either vehicle or PWCG at doses of 50 100 and200mgsdotkgminus1 BW for 10 weeks (119873 = 8group) 119875 value lt 001 and 0001 respectively compared with control group lowast lowastlowast lowastlowastlowast119875 valuelt 005 001 and 0001 respectively compared with diabetic rats which received vehicle (DM + vehicle)

Time durationOpacity index of the lens (mean plusmn SEM)

Control DM + vehicle DM + PWCG(50mgkgsdotBWminus1)

DM + PWCG(100mgkgsdotBWminus1)

DM + PWCG(200mgkgsdotBWminus1)

Baseline 000 plusmn 000 000 plusmn 000 000 plusmn 000 000 plusmn 000 000 plusmn 000Week 1 000 plusmn 000 000 plusmn 000 000 plusmn 000 000 plusmn 000 000 plusmn 000Week 2 000 plusmn 000 006 plusmn 006 000 plusmn 000 000 plusmn 000 000 plusmn 000Week 3 000 plusmn 000 050 plusmn 016 014 plusmn 010lowast 050 plusmn 013 022 plusmn 010Week 4 000 plusmn 000 100 plusmn 020 079 plusmn 024 100 plusmn 018 072 plusmn 023Week 5 000 plusmn 000 181 plusmn 031 100 plusmn 021lowast 125 plusmn 023 100 plusmn 028lowast

Week 6 008 plusmn 008 225 plusmn 032 150 plusmn 029 150 plusmn 022 111 plusmn 031lowastlowast

Week 7 017 plusmn 011 250 plusmn 034 171 plusmn 029 188 plusmn 029 144 plusmn 041lowast

Week 8 025 plusmn 013 306 plusmn 023 200 plusmn 030lowast 219 plusmn 025lowast 167 plusmn 040lowastlowastlowast

Week 9 050 plusmn 015 381 plusmn 010 229 plusmn 035lowastlowastlowast 238 plusmn 026lowastlowastlowast 178 plusmn 039lowastlowastlowast

Week 10 050 plusmn 015 400 plusmn 000 221 plusmn 037lowastlowastlowast 263 plusmn 026lowastlowastlowast 189 plusmn 040lowastlowastlowast

000

10000

20000

30000

40000

50000

60000

Bloo

d gl

ucos

e lev

el (m

gD

L)

ControlDM + vehicle

Time duration

Week 1 Week 5 Week 10

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 1 The average fasting blood glucose levels of all the experi-mental groups at 1-week 5-week and 10-week intervention period(119873 = 8group) 119875 value lt 0001 compared with control group

period PWCGat all doses used in this study failed to decreasethe fasting blood glucose level in diabetic rats

32 Anticataract Effect of PWCG Theeffect of PWCGon lensopacity was shown in Figure 2The lens opacity in control ratsalso developed cataract during 6ndash10-week period At 10-weekstudy period all lens from both right (1A and 1B) and left eyes(1a and 1b) of control rats were clear and normal Both right(2A and 2B) and left eyes (2a and 2b) of diabetic rats whichreceived vehicle showed lens opacity However the increasedlens opacity induced by diabetic condition was attenuated indiabetic rats which received all doses of PWCG (3Andash5A 3Bndash5B 3andash5a and 3bndash5b)

Table 1 showed that the lens opacity indices of diabeticrats which received vehicle started to increase at 3-week

treatment duration (119875 value lt 001 compared with controlrats) This change was still observed until the end of study(119875 value lt 0001 compared with control rats) Diabetic ratswhich received PWCG at dose of 200mgsdotkgminus1 significantlymitigated the elevation of lens opacity index induced by dia-betes at 5-week treatment (119875 valuelt 005 comparedwith dia-betic rats which received vehicle)This change persisted untilthe end of study (119875 value lt 001 005 0001 0001 0001 respcomparedwith diabetic rats which received vehicle) Diabeticrats which received PWCG at low dose (50mgsdotkgminus1 BW)also mitigated the elevation of lens opacity index induced bydiabetes but the significant changes were observed at 3-week5 week 8-week 9-week and 10-week treatment duration (119875value lt 005 005 005 0001 and 0001 resp compared withdiabetic rats which received vehicle) However the mediumdose of PWCG produced the significant mitigation effect onlens opacity index induced by diabetes only at 8-week 9-week and 10-week treatment duration (119875 value lt 005 0001and 0001 resp compared with diabetic rats which receivedvehicle)The effects of PWCGon lens opacity were confirmedby histopathological analysis of lens as shown in Figure 3It was found that at 10-week study period the lens capsulethickness and the density of epithelial cells of diabetic ratsubjected to vehicle treatment were decreased

In addition the reduction of differentiated lens fibersvacuoles and homogenized area was also observed (Figure3(b)) Diabetic rats which received PWCG at doses of 50100 and 200mgsdotkgminus1 BW showed the increased density ofepithelial cells and differentiated lens fiber thickness togetherwith the decreased vacuoles and homogenized area (Figures3(c) 3(d) and 3(e))

Figure 4 showed the effect of PWCG on the severity oflens damage Our results showed that the diabetic rats whichreceived vehicle showed the increased severity of cataract(119875 value lt 0001 compared with control rats) All doses ofPWCG significantly mitigated the enhanced cataract severityinduced by diabetes mellitus (119875 value lt 0001 all comparedwith diabetic rats which received vehicle)

Oxidative Medicine and Cellular Longevity 5

(1A) (2A) (3A) (4A) (5A)

(1B) (2B) (3B) (4B) (5B)

(1a) (2a) (3a) (4a) (5a)

(1b) (2b) (3b) (4b) (5b)

Figure 2 Representative photographs showing the effects of 10-week treatment of PWCG in prevention of cataract A photographs via slitlamp B photographs via camera 1 control rat 2 diabetic rat which received vehicle (DM + vehicle) 3 diabetic rat which received PWCGat dose of 50mgsdotkgminus1 BW (DM + PWCG 50mgsdotkgminus1 BW) 4 diabetic rat which received PWCG at dose of 100mgsdotkgminus1 BW (DM + PWCG100mgsdotkgminus1 BW) 5 diabetic rat which received PWCG at dose of 200mgsdotkgminus1 BW (DM + PWCG 200mgsdotkgminus1 BW)

33 The Effect of PWCG on Retinopathy in Diabetic Rats Theeffect of PWCG on histopathological change of retina wasshown in Figure 5 At 10-week study period the total retinalthickness (TRT) the thickness of the retinal outer nuclearlayer (ROT) and the number of cells in the ganglion celllayer (NG) of diabetic ratswhich received vehicle significantlydecreasedwhen compared to controlHowever the decreasedTRT ROT and NG induced by diabetic condition wereattenuated in diabetic rats which received all doses of PWCGFigures 6 7 and 8 showed that diabetic rats which receivedPWCG at doses of 50 100 and 200mgsdotkgminus1 BW significantlyincreased the TRT (119875 value lt 0001 all compared withdiabetic rats which received vehicle) ROT (119875 value lt 0001all compared with diabetic rats which received vehicle) andNG of retina (119875 value lt 0001 005 and 0001 resp comparedwith diabetic rats which received vehicle)

34 Effect of PWCG on Oxidative Stress Markers Based onthe crucial role of oxidative stress on cataract and retinopathythe effects of PWCG on oxidative stress markers includinglevel of MDA and the activities of SOD CAT and GPx inlens were carried out The results were shown in Figures 9

10 11 and 12 It was found that diabetic rats which receivedvehicle significantly increased MDA level (119875 value lt 0001compared with control) but decreased SOD CAT and GPxactivities (119875 value lt 0001 all compared with control) in lensPWCG at doses of 50 100 and 200mgsdotkgminus1 BW significantlymitigated the elevation of MDA level induced by diabetesmellitus (119875 value lt 0001 all compared with diabetic ratswhich received vehicle) Low dose of PWCG also mitigatedthe reduction of GPx activity in lens of diabetic rats (119875value lt 005 compared with diabetic rats which receivedvehicle) whereas high dose of PWCGmitigated the reductionof both CAT and GPx activities of lens of diabetic rats (119875value lt 001 and 0001 resp compared with diabetic ratswhich received vehicle) However no significant changes ofscavenger enzymes activities were observed in the diabeticrats which received medium dose of PWCG

35 Effect of PWCG on Aldose Reductase Activity In additionto the oxidative stress the polyol pathway also plays thecrucial role on the cataractogenesis Therefore the effectof PWCG on aldose reductase enzyme activity in lenswas also investigated and data were shown in Figure 13 It

6 Oxidative Medicine and Cellular Longevity

50 120583m

(a)

50 120583m

(b)

50 120583m

(c)

50 120583m

(d)

50 120583m

(e)

Figure 3 Photographs of transverse sections of eye balls (5 120583m thickness) stained with HampE which determined the cataract severity at theend of experiment using light microscope (week 10) (a) Control (b) DM + Vehicle (c) DM + PWCG (50mgkgsdotBW) (d) DM + PWCG(100mgkgsdotBW) and (e) DM + PWCG (200mgkgsdotBW)

000

050

100

150

200

250

300

350

400

Gra

de o

f cat

arac

t sev

erity

lowastlowastlowast

lowastlowastlowast

lowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 4 The cataract severity of control and diabetic rats (DM)which received either vehicle or PWCG at doses of 50 100 and200mgsdotkgminus1 BW for 10 weeks

119875 value lt 0001 compared withcontrol and lowastlowastlowast119875 value lt 0001 compared with vehicle

was demonstrated that aldose reductase activity in lens ofdiabetic rats was increased (119875 value lt 005 compared withcontrol) PWCG at dose of 200mgsdotkgminus1 BW could signifi-cantly decrease the elevation of aldose reductase induced bydiabetes mellitus in rat lens No other significant changeswere observed

4 Discussion

Diabetic cataract and retinopathy are the important causes ofblindness so the prevention of these conditions is very muchimportant Our results clearly demonstrated that PWCG sig-nificantly mitigated the reduction of ganglia in outer nuclearlayer (ONL) OUL thickness and total retina thickness Inaddition PWCG also showed anticataract effect Togetherwith the changes mentioned earlier the decreased oxidativestress and aldose reductase activity in rat lens were alsoobserved

It has been reported that the neurodegeneration of gan-glion cells plays the crucial role on the reduction of OULand total retina thickness [21] The degeneration of retinalganglion cells is under the influence of many factors suchas inflammation oxidative stress or exposure to advancedglycation end products [22] and polyol pathway [23] Ourdata also demonstrates the increased oxidative stress markerssuch asMDA level and the enhanced aldose reductase activityin lens of diabetic rats These changes are in agreement withthe previous study [22 23] Therefore we suggested thatthe effect of PWCG to attenuate retinopathy induced bydiabetes mellitus might be partly related to the decreasedoxidative stress and aldose reductase activity in rat lens Itwas found that the enhanced of GPx activity in rat lens mightbe responsible in part for the reduction of oxidative stress inrat lens of diabetic rats especially at low and high doses ofPWCG In addition to the increased GPx activity the highdose of PWCG also enhanced CAT in rat lens Therefore

Oxidative Medicine and Cellular Longevity 7

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(a)

50 120583m

RODS and CONES

ONL

OPLINL

IPLGCL

(b)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(c)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(d)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(e)

Figure 5 Light microscope of transverse section of eye ball (5 120583m thickness) stained with HampE which determined the pathological changeof retina at the end of experiment (week 10) (a) Control rat (b) diabetic rat which received vehicle (DM + vehicle) (c) diabetic rat whichreceived PWCG at dose of 50mgsdotkgminus1 BW (DM + PWCG 50mgsdotkgminus1 BW) (d) diabetic rat which received PWCG at dose of 100mgsdotkgminus1 BW(DM + PWCG 100mgsdotkgminus1 BW) (e) diabetic rat which received PWCG at dose of 200mgsdotkgminus1 BW (DM + PWCG 200mgsdotkgminus1 BW)

the elevation of both CAT and GPx might be responsible forthe decreased oxidative stress which in turn enhanced thesurvival of ganglion cells in OUL leading to the increasedtotal retina thickness However the decreased oxidative stressin rat lens induced by medium dose of PWCG might beassociated with other mechanism in addition to the elevationof the scavenger enzymes activities Since the excess oxidativestress occurs as the result of imbalance between oxidativestress formation and oxidative stress buffering capacities wesuggested that the decreased oxidative stress formationmightplay a role on the reduction of oxidative stress induced by thePWCG especially at the medium concentration

Oxidative stress and aldose reductase also play the crucialrole on the cataractogenesis in diabetic rats [24 25] In addi-tion it has been reported that antioxidant and aldose reduc-tase inhibitory effects can improve cataract [26] Thus theanticataractogenis of PWCG might also be associated withboth effects mentioned earlier Aldose reductase also playsa role on the pathophysiology of diabetic retinopathy [27]

Based on the basis of this information we do suggest thatthe effect of PWCG to improve retinopathy especially at highdose should also relate with the aldose reductase inhibitoryeffect

Previous study has demonstrated that flavonoids canprotect against oxidative stress and increase the survivalof ganglion cells [28] and prevent cataract [29] in diabeticrats Recent study has demonstrated that anthocyanins andanthocyanins-rich substance have the potential to protectagainst retinopathy [30] and diabetic cataract [26] In addi-tion ginger and its constituents also show the potential toprotect against diabetic retinopathy [31] Therefore the anti-cataractogenesis and antiretinopathy of PWCGmay be due tothe flavonoids such as anthocyanins in PWCG Other typesof flavonoidmay also play a role However further researchesare required to confirm the possible active ingredient

In this study the slit lamp examination clearly providedthe changes of lens characteristic However no pupil wasdilated during the grading of severity as that was performed

8 Oxidative Medicine and Cellular Longevity

0002000400060008000

100001200014000160001800020000

lowastlowastlowastlowastlowastlowast

lowastlowastlowast

Tota

l ret

inal

thic

knes

s (120583

m)

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 6 The total retinal thickness of retina (TRT) of control anddiabetic rats (DM) which received either vehicle or the combinationextract of purple waxy corn and ginger (PWCG) at doses of 50 100and 200mgsdotkgminus1 BW for 10weeks 119875 valuelt 0001 comparedwithcontrol and lowastlowastlowast119875 value lt 0001 compared with vehicle

000500

100015002000250030003500400045005000

The r

etin

al n

ucle

ar la

yer t

hick

ness

(120583m

)

lowastlowastlowast lowastlowastlowastlowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 7 The thickness of the retinal outer nuclear layer (ROT)of control and diabetic rats (DM) which received either vehicle orthe combination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks

119875 valuelt 0001 compared with control and lowastlowastlowast119875 value lt 0001 comparedwith vehicle

during the slit lamp examination in humanThismay possiblyinfluence the precision of grading scores which reflect thereal severity of cataract To obtain the precise severity ofcataract the slit lamp examination should be performedwhile pupil was dilated However all rats were assessed underthe condition without the pupil dilator so no confoundingerror about the beneficial effect of PWCG on cataract wasachieved

The lenticular opacification in control rats was alsoobserved during the 6ndash10 weeks of study period Based onthe previous study that stress hormone or glucocorticoids

000500

100015002000250030003500400045005000

The n

umbe

r of n

euro

nin

gan

glio

n ce

ll lay

er in

retin

a

lowastlowastlowastlowastlowastlowast

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 8 The number of cells in the ganglion cell layer (NG)in retina of control and diabetic rats (DM) which received eithervehicle or the combination extract of purple waxy corn and ginger(PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks 119875value lt 0001 compared with control and lowast lowastlowastlowast119875 value lt 005 and0001 compared with vehicle

MD

A (U

mg

prot

ein)

000

100

200

300

400

500

600

lowastlowastlowast lowastlowastlowastlowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 9 The level of malondialdehyde (MDA) (Umg protein) inlens of control and diabetic rats (DM) which received either vehicleor the combination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 = 8group)119875 value lt 0001 compared with control and lowastlowastlowast119875 value lt 0001

compared with diabetic rats which received vehicle (DM + vehicle)

can also induce cataract [32 33] we do suggest that thedevelopment of cataract in control group may possibly occuras the exposure to stress condition leading to the elevationof glucocorticoids hormone resulting in the cataractogenesisUnfortunately the level of corticosterone has not been deter-mined in this study Therefore the precise understandingmechanism of this change requires further investigation Inaddition spontaneous age-related cataract can also occurin laboratory rodents The prevalence is varied dependingon species Even young adolescent rodents can be found tohave cataract approximately 20 [34]Therefore the cataract

Oxidative Medicine and Cellular Longevity 9SO

D (U

mg

prot

ein)

000050100150200250300350400450

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 10 The activity of superoxide dismutase (SOD) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BWfor 10weeks(119873 = 8group) 119875 value lt 0001 compared with control

CAT

(Um

g pr

otei

n)

000

1000

2000

3000

4000

5000

6000

lowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 11 The activity of catalase (CAT) (Umg protein) in lensof control and diabetic rats (DM) which received either vehicle orthe combination extract of purple waxy corn and ginger (PWCG) atdoses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 = 8group)119875 value lt 0001 compared with control and lowastlowast119875 value lt 001

compared with diabetic rats which received vehicle (DM + vehicle)

observed in normal rats might occur either via stress orvia spontaneous cataractogenesis of the laboratory rodentsmentioned earlier

5 Conclusion

The combination extract of purple waxy corn seeds andginger rhizome (PWCG) can protect against diabetic cataractand diabetic retinopathyThepossible underlyingmechanismmay occur partly via the decreased oxidative stress and thesuppression of aldose reductase activity in lens The possibleactive ingredient may be associated with flavonoids such as

000020040060080100120140160180200

GPx

(Um

g pr

otei

n)

lowastlowastlowast

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 12 The activity of glutathione peroxidase (GPx) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10weeks (119873 = 8group) 119875 value lt 0001 compared with controland lowast lowastlowastlowast119875 value lt 005 0001 respectively compared with diabeticrats which received vehicle (DM + vehicle)

AR

activ

ity (n

mol

min

mg)

00000

00010

00020

00030

00040

00050

00060

00070

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 13 The activity of aldose reductase (AR) (nmolminmg)in lens of control and diabetic rats (DM) which received eithervehicle or the combination extract of purple waxy corn and ginger(PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 =8group) 119875 value lt 005 compared with control and lowast119875 value lt005 compared with diabetic rats which received vehicle (DM +vehicle)

anthocyanin and other flavonoids Further researches are stillessential to understand the precise underlying mechanismand possible active ingredient

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

10 Oxidative Medicine and Cellular Longevity

Acknowledgments

This study was supported by the Higher Education ResearchPromotion andNational Research University Project ofThai-land Office of the Higher Education Commission throughthe Food and Functional Food Research Cluster of KhonKaen University and the Integrative Complementary Alter-native Medicine Research and Development Center KhonKaen University Thailand Great thanks were also extendedto Research and Development Center of Herbal HealthProduct for providing the opportunity to collaborate in thesubcluster

References

[1] R L Engerman and T S Kern ldquoHyperglycemia as a cause ofdiabetic retinopathyrdquo Metabolism vol 35 supplement 1 no 4pp 20ndash23 1986

[2] A Pollreisz and U Schmidt-Erfurth ldquoDiabetic cataractmdashpathogenesis epidemiology and treatmentrdquo Journal of Ophthal-mology vol 2010 Article ID 608751 8 pages 2010

[3] I G Obrosova S S M Chung and P F Kador ldquoDiabeticcataracts mechanisms and managementrdquo DiabetesMetabolismResearch and Reviews vol 26 no 3 pp 172ndash180 2010

[4] J Ding and T Y Wong ldquoCurrent epidemiology of diabeticretinopathy and diabetic macular edemardquo Current DiabetesReports vol 12 no 4 pp 346ndash354 2012

[5] A Y W Lee and S S M Chung ldquoContributions of polyolpathway to oxidative stress in diabetic cataractrdquo The FASEBJournal vol 13 no 1 pp 23ndash30 1999

[6] S A Madsen-Bouterse and R A Kowluru ldquoOxidative stressand diabetic retinopathy pathophysiological mechanisms andtreatment perspectivesrdquo Reviews in Endocrine and MetabolicDisorders vol 9 no 4 pp 315ndash327 2008

[7] I G Obrosova and P F Kador ldquoAldose reductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 1389ndash2010 2011

[8] V R Drel P Pacher T K Ali et al ldquoAldose reductase inhibitorfidarestat counteracts diabetes-associated cataract formationretinal oxidative-nitrosative stress glial activation and apopto-sisrdquo International Journal of Molecular Medicine vol 21 no 6pp 667ndash676 2008

[9] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (Purple Waxy Corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[10] Y Morimitsu K Kubota T Tashiro E Hashizume T Kamiyaand T Osawa ldquoInhibitory effect of anthocyanins and coloredrice ondiabetic cataract formation in the rat lensesrdquo Interna-tional Congress Series vol 1245 pp 503ndash508 2002

[11] M Saraswat P Suryanarayana P Y Reddy M A Patil NBalakrishna and G B Reddy ldquoAntiglycating potential of Zin-giber officinalis and delay of diabetic cataract in ratsrdquoMolecularVision vol 16 pp 1525ndash1537 2010

[12] Y Li V H Tran C C Duke and B D Roufogalis ldquoPreventiveand protective properties of Zingiber officinale (Ginger) indiabetes mellitus diabetic complications and associated lipidand other metabolic disorders a brief reviewrdquo Evidence-basedComplementary and Alternative Medicine vol 2012 Article ID516870 10 pages 2012

[13] P Suryanarayana M Saraswat T Mrudula T P Krishna KKrishnaswamy andG B Reddy ldquoCurcumin and turmeric delaystreptozotocin-induced diabetic cataract in ratsrdquo InvestigativeOphthalmology amp Visual Science vol 46 no 6 pp 2092ndash20992005

[14] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[15] Y Sun L W Oberley and Y Li ldquoA simple method for clinicalassay of superoxide dismutaserdquo Clinical Chemistry vol 34 no3 pp 497ndash500 1988

[16] L Goth ldquoA simple method for determination of serum catalaseactivity and revision of reference rangerdquo Clinica Chimica Actavol 196 no 2-3 pp 143ndash151 1991

[17] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973

[18] M B Patel and S M Mishra ldquoAldose reductase inhibitoryactivity and anti catraract potential of some traditionallyacclaimed antidiabetic medicinal plantsrdquo Oriental Pharmacyand Experimental Medicine vol 9 no 3 pp 245ndash251 2009

[19] R Agarwal I Iezhitsa N A Awaludin et al ldquoEffects ofmagnesium taurate on the onset and progression of galactose-induced experimental cataract in vivo and in vitro evaluationrdquoExperimental Eye Research vol 110 pp 35ndash43 2013

[20] P M Martin P Roon T K van Ells V Ganapathy and SB Smith ldquoDeath of retinal neurons in streptozotocin-induceddiabetic micerdquo Investigative Ophthalmology and Visual Sciencevol 45 no 9 pp 3330ndash3336 2004

[21] H W van Dijk F D Verbraak P H B Kok et al ldquoDecreasedretinal ganglion cell layer thickness in patients with type 1diabetesrdquo Investigative Ophthalmology and Visual Science vol51 no 7 pp 3660ndash3665 2010

[22] T S Kern and A J Barber ldquoRetinal ganglion cells in diabetesrdquoJournal of Physiology vol 586 no 18 pp 4401ndash4408 2008

[23] G A Ramadan ldquoSorbitol-induced diabetic-like retinal lesionsin rats microscopic studyrdquo American Journal of Pharmacologyand Toxicology vol 2 no 2 pp 89ndash97 2007

[24] B Ozmen D Ozmen E Erkin I Guner S Habif and OBaymdir ldquoLens superoxide dismutase and catalase activities indiabetic cataractrdquoClinical Biochemistry vol 35 no 1 pp 69ndash722002

[25] A B M Reddy R Tammali R Mishra S Srivastava S KSrivastava and K V Ramana ldquoAldose reductase deficiencyprotects sugar-induced lens opacification in ratsrdquo Chemico-Biological Interactions vol 191 no 1ndash3 pp 346ndash350 2011

[26] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (purple waxy corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[27] I G Obrosova and P F Kador ldquoAldosereductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 373ndash385 2011

[28] PMaher andAHanneken ldquoFlavonoids protect retinal ganglioncells from oxidative stress-induced deathrdquo Investigative Oph-thalmology and Visual Science vol 46 no 12 pp 4796ndash48032005

[29] M Stefek ldquoNatural flavonoids as potential multifunctionalagents in prevention of diabetic cataractrdquo InterdisciplinaryToxicology vol 4 no 2 pp 69ndash77 2011

Oxidative Medicine and Cellular Longevity 11

[30] S F Nabavi S Habtemariam M Daglia N Shafighi A JBarber and S M Nabavi ldquoAnthocyanins as a potential therapyfor diabetic retinopathyrdquo Current Medicinal Chemistry vol 22no 1 pp 51ndash58 2014

[31] A H Rahmani F M Al Shabrmi and S M Aly ldquoActiveingredients of ginger as potential candidates in the preventionand treatment of diseases via modulation of biological activ-itiesrdquo International Journal of Physiology Pathophysiology andPharmacology vol 6 no 2 pp 125ndash136 2014

[32] H Watanabe H Kosano and H Nishigori ldquoSteroid-inducedshort term diabetes in chick embryo reversible effects of insulinon metabolic changes and cataract formationrdquo InvestigativeOphthalmology amp Visual Science vol 41 no 7 pp 1846ndash18522000

[33] E R James ldquoThe etiology of steroid cataractrdquo Journal of OcularPharmacology and Therapeutics vol 23 no 5 pp 403ndash4202007

[34] P Greaves Histopathological of Preclinical Toxicity StudiesElsevier New York NY USA 3rd edition 2007

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

2 Oxidative Medicine and Cellular Longevity

pilot data show that the antioxidant effect of the combinationof purple waxy corn and ginger (PWCG) is more potent thanthe antioxidant effect of either purple waxy corn or gingeralone (pilot data) However all of the evidence mentionedearlier is the in vitro studyTherefore we aimed to determinethe in vivo effect of PWCG on cataract retinopathy oxidativestress and aldose reductase in lens of diabetic rats

2 Materials and Methods

21 Plant Material and Extract Preparation The plant mate-rials used in this study were dried seeds of purple waxy cornor Zea mays L (purple color KKU open pollinated cultivar)and rhizomes of ginger or Zingiber officinale Roscoe Theywere harvested during September 2012 and authenticated byAssociate Kamol Lertrat and Dr Bhalang Suriharn Facultyof Agriculture Khon Kaen University Khon Kaen ThailandThe voucher specimens (voucher specimen 2012001 and2012002) were kept at the Integrative Complementary Alter-native Medicine Research and Development Center KhonKaen University The dried seeds of purple waxy corn andrhizomes of ginger were extracted with 50 hydroalcoholicsolvent by maceration method at a ratio of 2 5 and 1 5(weight volume) respectively The samples were maceratedat room temperature for 3 days Both of the yielded extractswere concentrated by lyophilization and kept at 4∘C forfurther study The percentage yields of purple waxy cornand ginger extracts were 572 and 2526 respectively Thecombination extract was prepared by mixing the purplewaxy corn and ginger extracts at a ratio of 1 4 (this ratioprovided the highest anticataract potential) The contentsof total phenolic compounds and anthocyaninsin the com-bination extract were 4482 plusmn 237mgL GAEmg extractand 39242plusmn003mgL cyanidin-3-glucoside equivalentsmgextract respectively

22 Experimental Design The experimental animals usedin this study were male Wistar rats at the weight of 200ndash250 g (119899 = 8 per group) The animals were maintainedand treated in accordance with the guideline and approvalof the Ethical Committee on Animals Experiments of KhonKaenUniversity (AEKKU982555) All rats were divided intovarious groups as follows

Group I Control group all rats in this group were adminis-tered citrate buffer a vehicle of streptozotocin (STZ)

Group II DM + vehicle group the animals in this groupwere induced diabetesmellitus via single injection of STZ andreceived vehicle of the extract or distilled water

Group IIIndashV The animals in this group had DM + thecombination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW respectively

All rats in groups IIndashV were induced diabetes cataractby a single injection of STZ which was dissolved in citratebuffer (pH 45) at dose of 55mgsdotkgminus1 BW The animalswhich showed the blood sugar levels more than 250mgsdotdLminus1were recruited for further study All rats were treated with

the assigned interventions once daily at 3 days after injectionof STZ and maintained for 10 weeks Lens opacity wasevaluated every week using slit lamp microscope At the endof study lens were collected and the level of malondialdehyde(MDA) and the activities of scavenging enzymes activitiesincluding superoxide dismutase (SOD) catalase (CAT) andglutathione peroxidase (GPx) in the lens together with lenshistomorphology were determined

23 Determination of Fasting Blood Glucose Level Afterfasting overnight blood was collected from rat tails andfasting blood glucose levels were determined using ACCU-CHEK active This process was performed at the first weekand every 5 weeks throughout the study period

24 Cataract Evaluation via Slit Lamp Rat eyes were eval-uated every week using slit lamp microscope (DIOPTRIX-HAWKEYE France) by trained observer who was blind tothe treatment The severity of cataract was graded as 5 stagesaccording to the method of Suryanarayana and coworkers[13] as follows

Stage 0 Lens were clear and no vacuoles were observed

Stage 1 Vacuoles cover approximately one-half of the surfacesof the anterior pole forming a subcapsular cataract

Stage 2 Some vacuoles have disappeared and the cortexexhibits a hazy opacity

Stage 3 A hazy cortex remains and dense nuclear opacity ispresent

Stage 4 A mature cataract is observed as a dense opacity inboth cortex and nucleus

Data were presented as opacity index which was calcu-lated from the following formula

Opacity index

=

(number of eyes in each stage times stage of the eye)total number of eyes within group

(1)

25 Homogenate Preparation At the end of experimentlens were collected and homogenized in 10wv of 01Mphosphate buffer pH 74 containing 1mM EDTA Then thehomogenate was centrifuged at 10000 g at 4∘C for 1 hour andthe supernatantwas separated and used for the determinationof biochemical parameters

26 Determination of Malondialdehyde (MDA) Level Levelof malondialdehyde (MDA) a lipid peroxidationmarker wasmonitored by using thiobarbituric acid reacting substances(TBARS) assay In brief 100 120583L of sample was mixed withthe solution containing 100120583L of 81 (wv) sodium dodecylsulphate 750120583L 20 (vv) acetic acid (pH 35) and 750 120583L of08 thiobarbituric acid (TBA) The solution was heated ina water bath at 95∘C for one hour and cooled immediately

Oxidative Medicine and Cellular Longevity 3

under running tap water Then 500120583L chilled water and2500 120583L of butanol and pyridine [15 1 vv] were added intoeach tube and mixed thoroughly with vortex Then thesolutionwas centrifuged at 800timesg for 20minTheupper layerwas separated and absorbance was measured at 532 nm 133-Tetra ethoxy propane (TEP) was used as the reference [14]The level of MDA was expressed as Umg protein

27 Superoxide Dismutase (SOD) Assay The determinationof SOD activity was carried out via nitro blue tetrazolium(NBT) reduction assay In this assay the xanthine-xanthineoxidase system was used as a superoxide generator In briefthe reaction mixture contained 20 120583L of sample and 200 120583Lof reaction mixture consisting of 57mM phosphate buffersolution (KH

2

PO4

) 01mM EDTA 10mM cytochrome Csolution and 50 120583M of xanthine solution and 20 120583L ofxanthine oxidase solution (090mUmL) were prepared at25∘C The optical density was measured at 415 nm A systemdevoid of enzyme served as the control and three parallelexperimentswere conducted [15] SODactivitywas expressedas Umg protein

28 Catalase (CAT) Assay Lens catalase activity was deter-mined based on the ability of the enzyme to break downH2

O2

In brief 10 120583L of sample was mixed with the reactionmixture containing 50 120583L of 30mM hydrogen peroxide (in50mMphosphate buffer pH 70) 25120583L of H

2

SO4

and 150 120583Lof KMnO

4

After mixing thoroughly the optical densitywas measured at 490 nm A system devoid of the substrate(hydrogen peroxide) served as the control The difference inabsorbance per unit time was expressed as the activity Theamount of enzyme which is required to decompose 10M ofhydrogen peroxide per minute at pH 70 and 25∘ is regardedas one unit [16] The value of CAT activity was expressed asUmg protein

29 Glutathione Peroxidase (GPx) Assay This assay was per-formed based on the glutathione recycling method by using551015840-dithiobis (2-nitrobenzoic acid) (DTNB) and glutathionereductase According to this method the reaction betweenDTNB and GSH gave rise to the generation of 2-nitro-5-thiobenzoic acid and GSSG Since 2-nitro-5-thiobenzoicacid was a yellow colored product GSH concentrationcould be determined by measuring absorbance at 412 nmIn brief a mixture containing a 20 120583L of sample and thereaction mixture consisting of 10 120583L of dithiothreitol (DTT)in 667mM potassium phosphate buffer (pH 7) 100120583L ofsodium azide in 667mM potassium phosphate buffer (pH7) 10 120583L of glutathione solution and 100 120583L of hydrogenperoxide was mixed thoroughly and incubated at roomtemperature for 5ndash10 minutes Then 10 120583L of DTNB (55-dithiobis-2-nitrobenzoic acid) was added and the opticaldensity at 412 nmwas recorded at 25∘Cover a period of 5minActivitieswere expressed as nmolesminmg lens protein [17]GPx activity was expressed as Umg protein

210 Aldose Reductase (AR) Activity Assay Aldose reductaseactivity was evaluated using spectrophotometric method

An assay mixture containing 07mL of phosphate buffer(0067mol) 01mL of NADPH (25 times 10minus5mol) 01mL of DL-glyceraldehyde (substrate 5 times 10minus4mol) and 01mL of lenssupernatant was prepared Absorbance was recorded againsta reference cuvette containing all other components exceptthe substrate DL-glyceraldehydeThe final pH of the reactionmixture was adjusted to pH = 62 The determination wasperformed after adding the substrate or DL-glyceraldehydeby measuring the decrease in NADPH absorbance at 390 nmover a 4-minute period [18] The enzyme activity wasexpressed as (nmolminmg)

211 Histopathological Analysis of Rat Lens and Retina Theeye balls from five rats per group were fixed in 10 formalinovernight embedded in paraffin sectioned at 5 120583m thick andstained with hematoxylin and eosin Light microscope wasused to evaluate the histomorphology of lens The severityof histomorphological change of lens was graded as a 5-grade score according to the method of Agarwal et al [19]as described as follow

Grade 0 Presence of anterior epithelium with lens fibers

Grade 1 Presence of anterior epithelium lens fibers andvacuoles

Grade 2 Presence of anterior epithelium lens fibers vacuolesand homogenized area

Grade 3 Absence of anterior epithelium presence of lensfibers vacuoles and homogenized area

Grade 4 Presence of lens fibers and homogenized area onlyHistomorphological changes of retina including the total

retinal thickness (from inner limiting membrane to Bruchrsquosmembrane) the thickness of the retinal outer nuclear layerand the number of cells in the ganglion cell layer were alsoperformed The average thickness of retina was evaluatedusing 3 adjacent fields and total five images in each group [20]The results were showed as mean plusmn SEM

212 Statistical Analysis All data were presented as mean plusmnstandard error mean (mean plusmn SEM) The analysis of datawas performed using one-way analysis of variance (ANOVA)followed by the post hoc test of LSD via SPSS version 15Statistical differences were considered at 119875 value lt 005

3 Results

31 Effect of PWCG on Fasting Blood Sugar Level The effectof PWCG on the average fasting blood glucose levels wasdetermined and the results were shown in Figure 1 Theresults showed that diabetic rats which received distilledwater or vehicle used in this study significantly enhanced theblood sugar levels throughout the study period (119875 value lt0001 all compared with control rats which received distilledwater) It was found that the blood sugar levels of all diabeticrats were more than 300mgsdotdLminus1 throughout the study

4 Oxidative Medicine and Cellular Longevity

Table 1 Opacity index of the lens of normal and diabetic rats (DM) which received either vehicle or PWCG at doses of 50 100 and200mgsdotkgminus1 BW for 10 weeks (119873 = 8group) 119875 value lt 001 and 0001 respectively compared with control group lowast lowastlowast lowastlowastlowast119875 valuelt 005 001 and 0001 respectively compared with diabetic rats which received vehicle (DM + vehicle)

Time durationOpacity index of the lens (mean plusmn SEM)

Control DM + vehicle DM + PWCG(50mgkgsdotBWminus1)

DM + PWCG(100mgkgsdotBWminus1)

DM + PWCG(200mgkgsdotBWminus1)

Baseline 000 plusmn 000 000 plusmn 000 000 plusmn 000 000 plusmn 000 000 plusmn 000Week 1 000 plusmn 000 000 plusmn 000 000 plusmn 000 000 plusmn 000 000 plusmn 000Week 2 000 plusmn 000 006 plusmn 006 000 plusmn 000 000 plusmn 000 000 plusmn 000Week 3 000 plusmn 000 050 plusmn 016 014 plusmn 010lowast 050 plusmn 013 022 plusmn 010Week 4 000 plusmn 000 100 plusmn 020 079 plusmn 024 100 plusmn 018 072 plusmn 023Week 5 000 plusmn 000 181 plusmn 031 100 plusmn 021lowast 125 plusmn 023 100 plusmn 028lowast

Week 6 008 plusmn 008 225 plusmn 032 150 plusmn 029 150 plusmn 022 111 plusmn 031lowastlowast

Week 7 017 plusmn 011 250 plusmn 034 171 plusmn 029 188 plusmn 029 144 plusmn 041lowast

Week 8 025 plusmn 013 306 plusmn 023 200 plusmn 030lowast 219 plusmn 025lowast 167 plusmn 040lowastlowastlowast

Week 9 050 plusmn 015 381 plusmn 010 229 plusmn 035lowastlowastlowast 238 plusmn 026lowastlowastlowast 178 plusmn 039lowastlowastlowast

Week 10 050 plusmn 015 400 plusmn 000 221 plusmn 037lowastlowastlowast 263 plusmn 026lowastlowastlowast 189 plusmn 040lowastlowastlowast

000

10000

20000

30000

40000

50000

60000

Bloo

d gl

ucos

e lev

el (m

gD

L)

ControlDM + vehicle

Time duration

Week 1 Week 5 Week 10

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 1 The average fasting blood glucose levels of all the experi-mental groups at 1-week 5-week and 10-week intervention period(119873 = 8group) 119875 value lt 0001 compared with control group

period PWCGat all doses used in this study failed to decreasethe fasting blood glucose level in diabetic rats

32 Anticataract Effect of PWCG Theeffect of PWCGon lensopacity was shown in Figure 2The lens opacity in control ratsalso developed cataract during 6ndash10-week period At 10-weekstudy period all lens from both right (1A and 1B) and left eyes(1a and 1b) of control rats were clear and normal Both right(2A and 2B) and left eyes (2a and 2b) of diabetic rats whichreceived vehicle showed lens opacity However the increasedlens opacity induced by diabetic condition was attenuated indiabetic rats which received all doses of PWCG (3Andash5A 3Bndash5B 3andash5a and 3bndash5b)

Table 1 showed that the lens opacity indices of diabeticrats which received vehicle started to increase at 3-week

treatment duration (119875 value lt 001 compared with controlrats) This change was still observed until the end of study(119875 value lt 0001 compared with control rats) Diabetic ratswhich received PWCG at dose of 200mgsdotkgminus1 significantlymitigated the elevation of lens opacity index induced by dia-betes at 5-week treatment (119875 valuelt 005 comparedwith dia-betic rats which received vehicle)This change persisted untilthe end of study (119875 value lt 001 005 0001 0001 0001 respcomparedwith diabetic rats which received vehicle) Diabeticrats which received PWCG at low dose (50mgsdotkgminus1 BW)also mitigated the elevation of lens opacity index induced bydiabetes but the significant changes were observed at 3-week5 week 8-week 9-week and 10-week treatment duration (119875value lt 005 005 005 0001 and 0001 resp compared withdiabetic rats which received vehicle) However the mediumdose of PWCG produced the significant mitigation effect onlens opacity index induced by diabetes only at 8-week 9-week and 10-week treatment duration (119875 value lt 005 0001and 0001 resp compared with diabetic rats which receivedvehicle)The effects of PWCGon lens opacity were confirmedby histopathological analysis of lens as shown in Figure 3It was found that at 10-week study period the lens capsulethickness and the density of epithelial cells of diabetic ratsubjected to vehicle treatment were decreased

In addition the reduction of differentiated lens fibersvacuoles and homogenized area was also observed (Figure3(b)) Diabetic rats which received PWCG at doses of 50100 and 200mgsdotkgminus1 BW showed the increased density ofepithelial cells and differentiated lens fiber thickness togetherwith the decreased vacuoles and homogenized area (Figures3(c) 3(d) and 3(e))

Figure 4 showed the effect of PWCG on the severity oflens damage Our results showed that the diabetic rats whichreceived vehicle showed the increased severity of cataract(119875 value lt 0001 compared with control rats) All doses ofPWCG significantly mitigated the enhanced cataract severityinduced by diabetes mellitus (119875 value lt 0001 all comparedwith diabetic rats which received vehicle)

Oxidative Medicine and Cellular Longevity 5

(1A) (2A) (3A) (4A) (5A)

(1B) (2B) (3B) (4B) (5B)

(1a) (2a) (3a) (4a) (5a)

(1b) (2b) (3b) (4b) (5b)

Figure 2 Representative photographs showing the effects of 10-week treatment of PWCG in prevention of cataract A photographs via slitlamp B photographs via camera 1 control rat 2 diabetic rat which received vehicle (DM + vehicle) 3 diabetic rat which received PWCGat dose of 50mgsdotkgminus1 BW (DM + PWCG 50mgsdotkgminus1 BW) 4 diabetic rat which received PWCG at dose of 100mgsdotkgminus1 BW (DM + PWCG100mgsdotkgminus1 BW) 5 diabetic rat which received PWCG at dose of 200mgsdotkgminus1 BW (DM + PWCG 200mgsdotkgminus1 BW)

33 The Effect of PWCG on Retinopathy in Diabetic Rats Theeffect of PWCG on histopathological change of retina wasshown in Figure 5 At 10-week study period the total retinalthickness (TRT) the thickness of the retinal outer nuclearlayer (ROT) and the number of cells in the ganglion celllayer (NG) of diabetic ratswhich received vehicle significantlydecreasedwhen compared to controlHowever the decreasedTRT ROT and NG induced by diabetic condition wereattenuated in diabetic rats which received all doses of PWCGFigures 6 7 and 8 showed that diabetic rats which receivedPWCG at doses of 50 100 and 200mgsdotkgminus1 BW significantlyincreased the TRT (119875 value lt 0001 all compared withdiabetic rats which received vehicle) ROT (119875 value lt 0001all compared with diabetic rats which received vehicle) andNG of retina (119875 value lt 0001 005 and 0001 resp comparedwith diabetic rats which received vehicle)

34 Effect of PWCG on Oxidative Stress Markers Based onthe crucial role of oxidative stress on cataract and retinopathythe effects of PWCG on oxidative stress markers includinglevel of MDA and the activities of SOD CAT and GPx inlens were carried out The results were shown in Figures 9

10 11 and 12 It was found that diabetic rats which receivedvehicle significantly increased MDA level (119875 value lt 0001compared with control) but decreased SOD CAT and GPxactivities (119875 value lt 0001 all compared with control) in lensPWCG at doses of 50 100 and 200mgsdotkgminus1 BW significantlymitigated the elevation of MDA level induced by diabetesmellitus (119875 value lt 0001 all compared with diabetic ratswhich received vehicle) Low dose of PWCG also mitigatedthe reduction of GPx activity in lens of diabetic rats (119875value lt 005 compared with diabetic rats which receivedvehicle) whereas high dose of PWCGmitigated the reductionof both CAT and GPx activities of lens of diabetic rats (119875value lt 001 and 0001 resp compared with diabetic ratswhich received vehicle) However no significant changes ofscavenger enzymes activities were observed in the diabeticrats which received medium dose of PWCG

35 Effect of PWCG on Aldose Reductase Activity In additionto the oxidative stress the polyol pathway also plays thecrucial role on the cataractogenesis Therefore the effectof PWCG on aldose reductase enzyme activity in lenswas also investigated and data were shown in Figure 13 It

6 Oxidative Medicine and Cellular Longevity

50 120583m

(a)

50 120583m

(b)

50 120583m

(c)

50 120583m

(d)

50 120583m

(e)

Figure 3 Photographs of transverse sections of eye balls (5 120583m thickness) stained with HampE which determined the cataract severity at theend of experiment using light microscope (week 10) (a) Control (b) DM + Vehicle (c) DM + PWCG (50mgkgsdotBW) (d) DM + PWCG(100mgkgsdotBW) and (e) DM + PWCG (200mgkgsdotBW)

000

050

100

150

200

250

300

350

400

Gra

de o

f cat

arac

t sev

erity

lowastlowastlowast

lowastlowastlowast

lowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 4 The cataract severity of control and diabetic rats (DM)which received either vehicle or PWCG at doses of 50 100 and200mgsdotkgminus1 BW for 10 weeks

119875 value lt 0001 compared withcontrol and lowastlowastlowast119875 value lt 0001 compared with vehicle

was demonstrated that aldose reductase activity in lens ofdiabetic rats was increased (119875 value lt 005 compared withcontrol) PWCG at dose of 200mgsdotkgminus1 BW could signifi-cantly decrease the elevation of aldose reductase induced bydiabetes mellitus in rat lens No other significant changeswere observed

4 Discussion

Diabetic cataract and retinopathy are the important causes ofblindness so the prevention of these conditions is very muchimportant Our results clearly demonstrated that PWCG sig-nificantly mitigated the reduction of ganglia in outer nuclearlayer (ONL) OUL thickness and total retina thickness Inaddition PWCG also showed anticataract effect Togetherwith the changes mentioned earlier the decreased oxidativestress and aldose reductase activity in rat lens were alsoobserved

It has been reported that the neurodegeneration of gan-glion cells plays the crucial role on the reduction of OULand total retina thickness [21] The degeneration of retinalganglion cells is under the influence of many factors suchas inflammation oxidative stress or exposure to advancedglycation end products [22] and polyol pathway [23] Ourdata also demonstrates the increased oxidative stress markerssuch asMDA level and the enhanced aldose reductase activityin lens of diabetic rats These changes are in agreement withthe previous study [22 23] Therefore we suggested thatthe effect of PWCG to attenuate retinopathy induced bydiabetes mellitus might be partly related to the decreasedoxidative stress and aldose reductase activity in rat lens Itwas found that the enhanced of GPx activity in rat lens mightbe responsible in part for the reduction of oxidative stress inrat lens of diabetic rats especially at low and high doses ofPWCG In addition to the increased GPx activity the highdose of PWCG also enhanced CAT in rat lens Therefore

Oxidative Medicine and Cellular Longevity 7

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(a)

50 120583m

RODS and CONES

ONL

OPLINL

IPLGCL

(b)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(c)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(d)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(e)

Figure 5 Light microscope of transverse section of eye ball (5 120583m thickness) stained with HampE which determined the pathological changeof retina at the end of experiment (week 10) (a) Control rat (b) diabetic rat which received vehicle (DM + vehicle) (c) diabetic rat whichreceived PWCG at dose of 50mgsdotkgminus1 BW (DM + PWCG 50mgsdotkgminus1 BW) (d) diabetic rat which received PWCG at dose of 100mgsdotkgminus1 BW(DM + PWCG 100mgsdotkgminus1 BW) (e) diabetic rat which received PWCG at dose of 200mgsdotkgminus1 BW (DM + PWCG 200mgsdotkgminus1 BW)

the elevation of both CAT and GPx might be responsible forthe decreased oxidative stress which in turn enhanced thesurvival of ganglion cells in OUL leading to the increasedtotal retina thickness However the decreased oxidative stressin rat lens induced by medium dose of PWCG might beassociated with other mechanism in addition to the elevationof the scavenger enzymes activities Since the excess oxidativestress occurs as the result of imbalance between oxidativestress formation and oxidative stress buffering capacities wesuggested that the decreased oxidative stress formationmightplay a role on the reduction of oxidative stress induced by thePWCG especially at the medium concentration

Oxidative stress and aldose reductase also play the crucialrole on the cataractogenesis in diabetic rats [24 25] In addi-tion it has been reported that antioxidant and aldose reduc-tase inhibitory effects can improve cataract [26] Thus theanticataractogenis of PWCG might also be associated withboth effects mentioned earlier Aldose reductase also playsa role on the pathophysiology of diabetic retinopathy [27]

Based on the basis of this information we do suggest thatthe effect of PWCG to improve retinopathy especially at highdose should also relate with the aldose reductase inhibitoryeffect

Previous study has demonstrated that flavonoids canprotect against oxidative stress and increase the survivalof ganglion cells [28] and prevent cataract [29] in diabeticrats Recent study has demonstrated that anthocyanins andanthocyanins-rich substance have the potential to protectagainst retinopathy [30] and diabetic cataract [26] In addi-tion ginger and its constituents also show the potential toprotect against diabetic retinopathy [31] Therefore the anti-cataractogenesis and antiretinopathy of PWCGmay be due tothe flavonoids such as anthocyanins in PWCG Other typesof flavonoidmay also play a role However further researchesare required to confirm the possible active ingredient

In this study the slit lamp examination clearly providedthe changes of lens characteristic However no pupil wasdilated during the grading of severity as that was performed

8 Oxidative Medicine and Cellular Longevity

0002000400060008000

100001200014000160001800020000

lowastlowastlowastlowastlowastlowast

lowastlowastlowast

Tota

l ret

inal

thic

knes

s (120583

m)

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 6 The total retinal thickness of retina (TRT) of control anddiabetic rats (DM) which received either vehicle or the combinationextract of purple waxy corn and ginger (PWCG) at doses of 50 100and 200mgsdotkgminus1 BW for 10weeks 119875 valuelt 0001 comparedwithcontrol and lowastlowastlowast119875 value lt 0001 compared with vehicle

000500

100015002000250030003500400045005000

The r

etin

al n

ucle

ar la

yer t

hick

ness

(120583m

)

lowastlowastlowast lowastlowastlowastlowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 7 The thickness of the retinal outer nuclear layer (ROT)of control and diabetic rats (DM) which received either vehicle orthe combination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks

119875 valuelt 0001 compared with control and lowastlowastlowast119875 value lt 0001 comparedwith vehicle

during the slit lamp examination in humanThismay possiblyinfluence the precision of grading scores which reflect thereal severity of cataract To obtain the precise severity ofcataract the slit lamp examination should be performedwhile pupil was dilated However all rats were assessed underthe condition without the pupil dilator so no confoundingerror about the beneficial effect of PWCG on cataract wasachieved

The lenticular opacification in control rats was alsoobserved during the 6ndash10 weeks of study period Based onthe previous study that stress hormone or glucocorticoids

000500

100015002000250030003500400045005000

The n

umbe

r of n

euro

nin

gan

glio

n ce

ll lay

er in

retin

a

lowastlowastlowastlowastlowastlowast

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 8 The number of cells in the ganglion cell layer (NG)in retina of control and diabetic rats (DM) which received eithervehicle or the combination extract of purple waxy corn and ginger(PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks 119875value lt 0001 compared with control and lowast lowastlowastlowast119875 value lt 005 and0001 compared with vehicle

MD

A (U

mg

prot

ein)

000

100

200

300

400

500

600

lowastlowastlowast lowastlowastlowastlowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 9 The level of malondialdehyde (MDA) (Umg protein) inlens of control and diabetic rats (DM) which received either vehicleor the combination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 = 8group)119875 value lt 0001 compared with control and lowastlowastlowast119875 value lt 0001

compared with diabetic rats which received vehicle (DM + vehicle)

can also induce cataract [32 33] we do suggest that thedevelopment of cataract in control group may possibly occuras the exposure to stress condition leading to the elevationof glucocorticoids hormone resulting in the cataractogenesisUnfortunately the level of corticosterone has not been deter-mined in this study Therefore the precise understandingmechanism of this change requires further investigation Inaddition spontaneous age-related cataract can also occurin laboratory rodents The prevalence is varied dependingon species Even young adolescent rodents can be found tohave cataract approximately 20 [34]Therefore the cataract

Oxidative Medicine and Cellular Longevity 9SO

D (U

mg

prot

ein)

000050100150200250300350400450

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 10 The activity of superoxide dismutase (SOD) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BWfor 10weeks(119873 = 8group) 119875 value lt 0001 compared with control

CAT

(Um

g pr

otei

n)

000

1000

2000

3000

4000

5000

6000

lowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 11 The activity of catalase (CAT) (Umg protein) in lensof control and diabetic rats (DM) which received either vehicle orthe combination extract of purple waxy corn and ginger (PWCG) atdoses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 = 8group)119875 value lt 0001 compared with control and lowastlowast119875 value lt 001

compared with diabetic rats which received vehicle (DM + vehicle)

observed in normal rats might occur either via stress orvia spontaneous cataractogenesis of the laboratory rodentsmentioned earlier

5 Conclusion

The combination extract of purple waxy corn seeds andginger rhizome (PWCG) can protect against diabetic cataractand diabetic retinopathyThepossible underlyingmechanismmay occur partly via the decreased oxidative stress and thesuppression of aldose reductase activity in lens The possibleactive ingredient may be associated with flavonoids such as

000020040060080100120140160180200

GPx

(Um

g pr

otei

n)

lowastlowastlowast

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 12 The activity of glutathione peroxidase (GPx) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10weeks (119873 = 8group) 119875 value lt 0001 compared with controland lowast lowastlowastlowast119875 value lt 005 0001 respectively compared with diabeticrats which received vehicle (DM + vehicle)

AR

activ

ity (n

mol

min

mg)

00000

00010

00020

00030

00040

00050

00060

00070

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 13 The activity of aldose reductase (AR) (nmolminmg)in lens of control and diabetic rats (DM) which received eithervehicle or the combination extract of purple waxy corn and ginger(PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 =8group) 119875 value lt 005 compared with control and lowast119875 value lt005 compared with diabetic rats which received vehicle (DM +vehicle)

anthocyanin and other flavonoids Further researches are stillessential to understand the precise underlying mechanismand possible active ingredient

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

10 Oxidative Medicine and Cellular Longevity

Acknowledgments

This study was supported by the Higher Education ResearchPromotion andNational Research University Project ofThai-land Office of the Higher Education Commission throughthe Food and Functional Food Research Cluster of KhonKaen University and the Integrative Complementary Alter-native Medicine Research and Development Center KhonKaen University Thailand Great thanks were also extendedto Research and Development Center of Herbal HealthProduct for providing the opportunity to collaborate in thesubcluster

References

[1] R L Engerman and T S Kern ldquoHyperglycemia as a cause ofdiabetic retinopathyrdquo Metabolism vol 35 supplement 1 no 4pp 20ndash23 1986

[2] A Pollreisz and U Schmidt-Erfurth ldquoDiabetic cataractmdashpathogenesis epidemiology and treatmentrdquo Journal of Ophthal-mology vol 2010 Article ID 608751 8 pages 2010

[3] I G Obrosova S S M Chung and P F Kador ldquoDiabeticcataracts mechanisms and managementrdquo DiabetesMetabolismResearch and Reviews vol 26 no 3 pp 172ndash180 2010

[4] J Ding and T Y Wong ldquoCurrent epidemiology of diabeticretinopathy and diabetic macular edemardquo Current DiabetesReports vol 12 no 4 pp 346ndash354 2012

[5] A Y W Lee and S S M Chung ldquoContributions of polyolpathway to oxidative stress in diabetic cataractrdquo The FASEBJournal vol 13 no 1 pp 23ndash30 1999

[6] S A Madsen-Bouterse and R A Kowluru ldquoOxidative stressand diabetic retinopathy pathophysiological mechanisms andtreatment perspectivesrdquo Reviews in Endocrine and MetabolicDisorders vol 9 no 4 pp 315ndash327 2008

[7] I G Obrosova and P F Kador ldquoAldose reductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 1389ndash2010 2011

[8] V R Drel P Pacher T K Ali et al ldquoAldose reductase inhibitorfidarestat counteracts diabetes-associated cataract formationretinal oxidative-nitrosative stress glial activation and apopto-sisrdquo International Journal of Molecular Medicine vol 21 no 6pp 667ndash676 2008

[9] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (Purple Waxy Corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[10] Y Morimitsu K Kubota T Tashiro E Hashizume T Kamiyaand T Osawa ldquoInhibitory effect of anthocyanins and coloredrice ondiabetic cataract formation in the rat lensesrdquo Interna-tional Congress Series vol 1245 pp 503ndash508 2002

[11] M Saraswat P Suryanarayana P Y Reddy M A Patil NBalakrishna and G B Reddy ldquoAntiglycating potential of Zin-giber officinalis and delay of diabetic cataract in ratsrdquoMolecularVision vol 16 pp 1525ndash1537 2010

[12] Y Li V H Tran C C Duke and B D Roufogalis ldquoPreventiveand protective properties of Zingiber officinale (Ginger) indiabetes mellitus diabetic complications and associated lipidand other metabolic disorders a brief reviewrdquo Evidence-basedComplementary and Alternative Medicine vol 2012 Article ID516870 10 pages 2012

[13] P Suryanarayana M Saraswat T Mrudula T P Krishna KKrishnaswamy andG B Reddy ldquoCurcumin and turmeric delaystreptozotocin-induced diabetic cataract in ratsrdquo InvestigativeOphthalmology amp Visual Science vol 46 no 6 pp 2092ndash20992005

[14] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[15] Y Sun L W Oberley and Y Li ldquoA simple method for clinicalassay of superoxide dismutaserdquo Clinical Chemistry vol 34 no3 pp 497ndash500 1988

[16] L Goth ldquoA simple method for determination of serum catalaseactivity and revision of reference rangerdquo Clinica Chimica Actavol 196 no 2-3 pp 143ndash151 1991

[17] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973

[18] M B Patel and S M Mishra ldquoAldose reductase inhibitoryactivity and anti catraract potential of some traditionallyacclaimed antidiabetic medicinal plantsrdquo Oriental Pharmacyand Experimental Medicine vol 9 no 3 pp 245ndash251 2009

[19] R Agarwal I Iezhitsa N A Awaludin et al ldquoEffects ofmagnesium taurate on the onset and progression of galactose-induced experimental cataract in vivo and in vitro evaluationrdquoExperimental Eye Research vol 110 pp 35ndash43 2013

[20] P M Martin P Roon T K van Ells V Ganapathy and SB Smith ldquoDeath of retinal neurons in streptozotocin-induceddiabetic micerdquo Investigative Ophthalmology and Visual Sciencevol 45 no 9 pp 3330ndash3336 2004

[21] H W van Dijk F D Verbraak P H B Kok et al ldquoDecreasedretinal ganglion cell layer thickness in patients with type 1diabetesrdquo Investigative Ophthalmology and Visual Science vol51 no 7 pp 3660ndash3665 2010

[22] T S Kern and A J Barber ldquoRetinal ganglion cells in diabetesrdquoJournal of Physiology vol 586 no 18 pp 4401ndash4408 2008

[23] G A Ramadan ldquoSorbitol-induced diabetic-like retinal lesionsin rats microscopic studyrdquo American Journal of Pharmacologyand Toxicology vol 2 no 2 pp 89ndash97 2007

[24] B Ozmen D Ozmen E Erkin I Guner S Habif and OBaymdir ldquoLens superoxide dismutase and catalase activities indiabetic cataractrdquoClinical Biochemistry vol 35 no 1 pp 69ndash722002

[25] A B M Reddy R Tammali R Mishra S Srivastava S KSrivastava and K V Ramana ldquoAldose reductase deficiencyprotects sugar-induced lens opacification in ratsrdquo Chemico-Biological Interactions vol 191 no 1ndash3 pp 346ndash350 2011

[26] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (purple waxy corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[27] I G Obrosova and P F Kador ldquoAldosereductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 373ndash385 2011

[28] PMaher andAHanneken ldquoFlavonoids protect retinal ganglioncells from oxidative stress-induced deathrdquo Investigative Oph-thalmology and Visual Science vol 46 no 12 pp 4796ndash48032005

[29] M Stefek ldquoNatural flavonoids as potential multifunctionalagents in prevention of diabetic cataractrdquo InterdisciplinaryToxicology vol 4 no 2 pp 69ndash77 2011

Oxidative Medicine and Cellular Longevity 11

[30] S F Nabavi S Habtemariam M Daglia N Shafighi A JBarber and S M Nabavi ldquoAnthocyanins as a potential therapyfor diabetic retinopathyrdquo Current Medicinal Chemistry vol 22no 1 pp 51ndash58 2014

[31] A H Rahmani F M Al Shabrmi and S M Aly ldquoActiveingredients of ginger as potential candidates in the preventionand treatment of diseases via modulation of biological activ-itiesrdquo International Journal of Physiology Pathophysiology andPharmacology vol 6 no 2 pp 125ndash136 2014

[32] H Watanabe H Kosano and H Nishigori ldquoSteroid-inducedshort term diabetes in chick embryo reversible effects of insulinon metabolic changes and cataract formationrdquo InvestigativeOphthalmology amp Visual Science vol 41 no 7 pp 1846ndash18522000

[33] E R James ldquoThe etiology of steroid cataractrdquo Journal of OcularPharmacology and Therapeutics vol 23 no 5 pp 403ndash4202007

[34] P Greaves Histopathological of Preclinical Toxicity StudiesElsevier New York NY USA 3rd edition 2007

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Oxidative Medicine and Cellular Longevity 3

under running tap water Then 500120583L chilled water and2500 120583L of butanol and pyridine [15 1 vv] were added intoeach tube and mixed thoroughly with vortex Then thesolutionwas centrifuged at 800timesg for 20minTheupper layerwas separated and absorbance was measured at 532 nm 133-Tetra ethoxy propane (TEP) was used as the reference [14]The level of MDA was expressed as Umg protein

27 Superoxide Dismutase (SOD) Assay The determinationof SOD activity was carried out via nitro blue tetrazolium(NBT) reduction assay In this assay the xanthine-xanthineoxidase system was used as a superoxide generator In briefthe reaction mixture contained 20 120583L of sample and 200 120583Lof reaction mixture consisting of 57mM phosphate buffersolution (KH

2

PO4

) 01mM EDTA 10mM cytochrome Csolution and 50 120583M of xanthine solution and 20 120583L ofxanthine oxidase solution (090mUmL) were prepared at25∘C The optical density was measured at 415 nm A systemdevoid of enzyme served as the control and three parallelexperimentswere conducted [15] SODactivitywas expressedas Umg protein

28 Catalase (CAT) Assay Lens catalase activity was deter-mined based on the ability of the enzyme to break downH2

O2

In brief 10 120583L of sample was mixed with the reactionmixture containing 50 120583L of 30mM hydrogen peroxide (in50mMphosphate buffer pH 70) 25120583L of H

2

SO4

and 150 120583Lof KMnO

4

After mixing thoroughly the optical densitywas measured at 490 nm A system devoid of the substrate(hydrogen peroxide) served as the control The difference inabsorbance per unit time was expressed as the activity Theamount of enzyme which is required to decompose 10M ofhydrogen peroxide per minute at pH 70 and 25∘ is regardedas one unit [16] The value of CAT activity was expressed asUmg protein

29 Glutathione Peroxidase (GPx) Assay This assay was per-formed based on the glutathione recycling method by using551015840-dithiobis (2-nitrobenzoic acid) (DTNB) and glutathionereductase According to this method the reaction betweenDTNB and GSH gave rise to the generation of 2-nitro-5-thiobenzoic acid and GSSG Since 2-nitro-5-thiobenzoicacid was a yellow colored product GSH concentrationcould be determined by measuring absorbance at 412 nmIn brief a mixture containing a 20 120583L of sample and thereaction mixture consisting of 10 120583L of dithiothreitol (DTT)in 667mM potassium phosphate buffer (pH 7) 100120583L ofsodium azide in 667mM potassium phosphate buffer (pH7) 10 120583L of glutathione solution and 100 120583L of hydrogenperoxide was mixed thoroughly and incubated at roomtemperature for 5ndash10 minutes Then 10 120583L of DTNB (55-dithiobis-2-nitrobenzoic acid) was added and the opticaldensity at 412 nmwas recorded at 25∘Cover a period of 5minActivitieswere expressed as nmolesminmg lens protein [17]GPx activity was expressed as Umg protein

210 Aldose Reductase (AR) Activity Assay Aldose reductaseactivity was evaluated using spectrophotometric method

An assay mixture containing 07mL of phosphate buffer(0067mol) 01mL of NADPH (25 times 10minus5mol) 01mL of DL-glyceraldehyde (substrate 5 times 10minus4mol) and 01mL of lenssupernatant was prepared Absorbance was recorded againsta reference cuvette containing all other components exceptthe substrate DL-glyceraldehydeThe final pH of the reactionmixture was adjusted to pH = 62 The determination wasperformed after adding the substrate or DL-glyceraldehydeby measuring the decrease in NADPH absorbance at 390 nmover a 4-minute period [18] The enzyme activity wasexpressed as (nmolminmg)

211 Histopathological Analysis of Rat Lens and Retina Theeye balls from five rats per group were fixed in 10 formalinovernight embedded in paraffin sectioned at 5 120583m thick andstained with hematoxylin and eosin Light microscope wasused to evaluate the histomorphology of lens The severityof histomorphological change of lens was graded as a 5-grade score according to the method of Agarwal et al [19]as described as follow

Grade 0 Presence of anterior epithelium with lens fibers

Grade 1 Presence of anterior epithelium lens fibers andvacuoles

Grade 2 Presence of anterior epithelium lens fibers vacuolesand homogenized area

Grade 3 Absence of anterior epithelium presence of lensfibers vacuoles and homogenized area

Grade 4 Presence of lens fibers and homogenized area onlyHistomorphological changes of retina including the total

retinal thickness (from inner limiting membrane to Bruchrsquosmembrane) the thickness of the retinal outer nuclear layerand the number of cells in the ganglion cell layer were alsoperformed The average thickness of retina was evaluatedusing 3 adjacent fields and total five images in each group [20]The results were showed as mean plusmn SEM

212 Statistical Analysis All data were presented as mean plusmnstandard error mean (mean plusmn SEM) The analysis of datawas performed using one-way analysis of variance (ANOVA)followed by the post hoc test of LSD via SPSS version 15Statistical differences were considered at 119875 value lt 005

3 Results

31 Effect of PWCG on Fasting Blood Sugar Level The effectof PWCG on the average fasting blood glucose levels wasdetermined and the results were shown in Figure 1 Theresults showed that diabetic rats which received distilledwater or vehicle used in this study significantly enhanced theblood sugar levels throughout the study period (119875 value lt0001 all compared with control rats which received distilledwater) It was found that the blood sugar levels of all diabeticrats were more than 300mgsdotdLminus1 throughout the study

4 Oxidative Medicine and Cellular Longevity

Table 1 Opacity index of the lens of normal and diabetic rats (DM) which received either vehicle or PWCG at doses of 50 100 and200mgsdotkgminus1 BW for 10 weeks (119873 = 8group) 119875 value lt 001 and 0001 respectively compared with control group lowast lowastlowast lowastlowastlowast119875 valuelt 005 001 and 0001 respectively compared with diabetic rats which received vehicle (DM + vehicle)

Time durationOpacity index of the lens (mean plusmn SEM)

Control DM + vehicle DM + PWCG(50mgkgsdotBWminus1)

DM + PWCG(100mgkgsdotBWminus1)

DM + PWCG(200mgkgsdotBWminus1)

Baseline 000 plusmn 000 000 plusmn 000 000 plusmn 000 000 plusmn 000 000 plusmn 000Week 1 000 plusmn 000 000 plusmn 000 000 plusmn 000 000 plusmn 000 000 plusmn 000Week 2 000 plusmn 000 006 plusmn 006 000 plusmn 000 000 plusmn 000 000 plusmn 000Week 3 000 plusmn 000 050 plusmn 016 014 plusmn 010lowast 050 plusmn 013 022 plusmn 010Week 4 000 plusmn 000 100 plusmn 020 079 plusmn 024 100 plusmn 018 072 plusmn 023Week 5 000 plusmn 000 181 plusmn 031 100 plusmn 021lowast 125 plusmn 023 100 plusmn 028lowast

Week 6 008 plusmn 008 225 plusmn 032 150 plusmn 029 150 plusmn 022 111 plusmn 031lowastlowast

Week 7 017 plusmn 011 250 plusmn 034 171 plusmn 029 188 plusmn 029 144 plusmn 041lowast

Week 8 025 plusmn 013 306 plusmn 023 200 plusmn 030lowast 219 plusmn 025lowast 167 plusmn 040lowastlowastlowast

Week 9 050 plusmn 015 381 plusmn 010 229 plusmn 035lowastlowastlowast 238 plusmn 026lowastlowastlowast 178 plusmn 039lowastlowastlowast

Week 10 050 plusmn 015 400 plusmn 000 221 plusmn 037lowastlowastlowast 263 plusmn 026lowastlowastlowast 189 plusmn 040lowastlowastlowast

000

10000

20000

30000

40000

50000

60000

Bloo

d gl

ucos

e lev

el (m

gD

L)

ControlDM + vehicle

Time duration

Week 1 Week 5 Week 10

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 1 The average fasting blood glucose levels of all the experi-mental groups at 1-week 5-week and 10-week intervention period(119873 = 8group) 119875 value lt 0001 compared with control group

period PWCGat all doses used in this study failed to decreasethe fasting blood glucose level in diabetic rats

32 Anticataract Effect of PWCG Theeffect of PWCGon lensopacity was shown in Figure 2The lens opacity in control ratsalso developed cataract during 6ndash10-week period At 10-weekstudy period all lens from both right (1A and 1B) and left eyes(1a and 1b) of control rats were clear and normal Both right(2A and 2B) and left eyes (2a and 2b) of diabetic rats whichreceived vehicle showed lens opacity However the increasedlens opacity induced by diabetic condition was attenuated indiabetic rats which received all doses of PWCG (3Andash5A 3Bndash5B 3andash5a and 3bndash5b)

Table 1 showed that the lens opacity indices of diabeticrats which received vehicle started to increase at 3-week

treatment duration (119875 value lt 001 compared with controlrats) This change was still observed until the end of study(119875 value lt 0001 compared with control rats) Diabetic ratswhich received PWCG at dose of 200mgsdotkgminus1 significantlymitigated the elevation of lens opacity index induced by dia-betes at 5-week treatment (119875 valuelt 005 comparedwith dia-betic rats which received vehicle)This change persisted untilthe end of study (119875 value lt 001 005 0001 0001 0001 respcomparedwith diabetic rats which received vehicle) Diabeticrats which received PWCG at low dose (50mgsdotkgminus1 BW)also mitigated the elevation of lens opacity index induced bydiabetes but the significant changes were observed at 3-week5 week 8-week 9-week and 10-week treatment duration (119875value lt 005 005 005 0001 and 0001 resp compared withdiabetic rats which received vehicle) However the mediumdose of PWCG produced the significant mitigation effect onlens opacity index induced by diabetes only at 8-week 9-week and 10-week treatment duration (119875 value lt 005 0001and 0001 resp compared with diabetic rats which receivedvehicle)The effects of PWCGon lens opacity were confirmedby histopathological analysis of lens as shown in Figure 3It was found that at 10-week study period the lens capsulethickness and the density of epithelial cells of diabetic ratsubjected to vehicle treatment were decreased

In addition the reduction of differentiated lens fibersvacuoles and homogenized area was also observed (Figure3(b)) Diabetic rats which received PWCG at doses of 50100 and 200mgsdotkgminus1 BW showed the increased density ofepithelial cells and differentiated lens fiber thickness togetherwith the decreased vacuoles and homogenized area (Figures3(c) 3(d) and 3(e))

Figure 4 showed the effect of PWCG on the severity oflens damage Our results showed that the diabetic rats whichreceived vehicle showed the increased severity of cataract(119875 value lt 0001 compared with control rats) All doses ofPWCG significantly mitigated the enhanced cataract severityinduced by diabetes mellitus (119875 value lt 0001 all comparedwith diabetic rats which received vehicle)

Oxidative Medicine and Cellular Longevity 5

(1A) (2A) (3A) (4A) (5A)

(1B) (2B) (3B) (4B) (5B)

(1a) (2a) (3a) (4a) (5a)

(1b) (2b) (3b) (4b) (5b)

Figure 2 Representative photographs showing the effects of 10-week treatment of PWCG in prevention of cataract A photographs via slitlamp B photographs via camera 1 control rat 2 diabetic rat which received vehicle (DM + vehicle) 3 diabetic rat which received PWCGat dose of 50mgsdotkgminus1 BW (DM + PWCG 50mgsdotkgminus1 BW) 4 diabetic rat which received PWCG at dose of 100mgsdotkgminus1 BW (DM + PWCG100mgsdotkgminus1 BW) 5 diabetic rat which received PWCG at dose of 200mgsdotkgminus1 BW (DM + PWCG 200mgsdotkgminus1 BW)

33 The Effect of PWCG on Retinopathy in Diabetic Rats Theeffect of PWCG on histopathological change of retina wasshown in Figure 5 At 10-week study period the total retinalthickness (TRT) the thickness of the retinal outer nuclearlayer (ROT) and the number of cells in the ganglion celllayer (NG) of diabetic ratswhich received vehicle significantlydecreasedwhen compared to controlHowever the decreasedTRT ROT and NG induced by diabetic condition wereattenuated in diabetic rats which received all doses of PWCGFigures 6 7 and 8 showed that diabetic rats which receivedPWCG at doses of 50 100 and 200mgsdotkgminus1 BW significantlyincreased the TRT (119875 value lt 0001 all compared withdiabetic rats which received vehicle) ROT (119875 value lt 0001all compared with diabetic rats which received vehicle) andNG of retina (119875 value lt 0001 005 and 0001 resp comparedwith diabetic rats which received vehicle)

34 Effect of PWCG on Oxidative Stress Markers Based onthe crucial role of oxidative stress on cataract and retinopathythe effects of PWCG on oxidative stress markers includinglevel of MDA and the activities of SOD CAT and GPx inlens were carried out The results were shown in Figures 9

10 11 and 12 It was found that diabetic rats which receivedvehicle significantly increased MDA level (119875 value lt 0001compared with control) but decreased SOD CAT and GPxactivities (119875 value lt 0001 all compared with control) in lensPWCG at doses of 50 100 and 200mgsdotkgminus1 BW significantlymitigated the elevation of MDA level induced by diabetesmellitus (119875 value lt 0001 all compared with diabetic ratswhich received vehicle) Low dose of PWCG also mitigatedthe reduction of GPx activity in lens of diabetic rats (119875value lt 005 compared with diabetic rats which receivedvehicle) whereas high dose of PWCGmitigated the reductionof both CAT and GPx activities of lens of diabetic rats (119875value lt 001 and 0001 resp compared with diabetic ratswhich received vehicle) However no significant changes ofscavenger enzymes activities were observed in the diabeticrats which received medium dose of PWCG

35 Effect of PWCG on Aldose Reductase Activity In additionto the oxidative stress the polyol pathway also plays thecrucial role on the cataractogenesis Therefore the effectof PWCG on aldose reductase enzyme activity in lenswas also investigated and data were shown in Figure 13 It

6 Oxidative Medicine and Cellular Longevity

50 120583m

(a)

50 120583m

(b)

50 120583m

(c)

50 120583m

(d)

50 120583m

(e)

Figure 3 Photographs of transverse sections of eye balls (5 120583m thickness) stained with HampE which determined the cataract severity at theend of experiment using light microscope (week 10) (a) Control (b) DM + Vehicle (c) DM + PWCG (50mgkgsdotBW) (d) DM + PWCG(100mgkgsdotBW) and (e) DM + PWCG (200mgkgsdotBW)

000

050

100

150

200

250

300

350

400

Gra

de o

f cat

arac

t sev

erity

lowastlowastlowast

lowastlowastlowast

lowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 4 The cataract severity of control and diabetic rats (DM)which received either vehicle or PWCG at doses of 50 100 and200mgsdotkgminus1 BW for 10 weeks

119875 value lt 0001 compared withcontrol and lowastlowastlowast119875 value lt 0001 compared with vehicle

was demonstrated that aldose reductase activity in lens ofdiabetic rats was increased (119875 value lt 005 compared withcontrol) PWCG at dose of 200mgsdotkgminus1 BW could signifi-cantly decrease the elevation of aldose reductase induced bydiabetes mellitus in rat lens No other significant changeswere observed

4 Discussion

Diabetic cataract and retinopathy are the important causes ofblindness so the prevention of these conditions is very muchimportant Our results clearly demonstrated that PWCG sig-nificantly mitigated the reduction of ganglia in outer nuclearlayer (ONL) OUL thickness and total retina thickness Inaddition PWCG also showed anticataract effect Togetherwith the changes mentioned earlier the decreased oxidativestress and aldose reductase activity in rat lens were alsoobserved

It has been reported that the neurodegeneration of gan-glion cells plays the crucial role on the reduction of OULand total retina thickness [21] The degeneration of retinalganglion cells is under the influence of many factors suchas inflammation oxidative stress or exposure to advancedglycation end products [22] and polyol pathway [23] Ourdata also demonstrates the increased oxidative stress markerssuch asMDA level and the enhanced aldose reductase activityin lens of diabetic rats These changes are in agreement withthe previous study [22 23] Therefore we suggested thatthe effect of PWCG to attenuate retinopathy induced bydiabetes mellitus might be partly related to the decreasedoxidative stress and aldose reductase activity in rat lens Itwas found that the enhanced of GPx activity in rat lens mightbe responsible in part for the reduction of oxidative stress inrat lens of diabetic rats especially at low and high doses ofPWCG In addition to the increased GPx activity the highdose of PWCG also enhanced CAT in rat lens Therefore

Oxidative Medicine and Cellular Longevity 7

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(a)

50 120583m

RODS and CONES

ONL

OPLINL

IPLGCL

(b)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(c)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(d)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(e)

Figure 5 Light microscope of transverse section of eye ball (5 120583m thickness) stained with HampE which determined the pathological changeof retina at the end of experiment (week 10) (a) Control rat (b) diabetic rat which received vehicle (DM + vehicle) (c) diabetic rat whichreceived PWCG at dose of 50mgsdotkgminus1 BW (DM + PWCG 50mgsdotkgminus1 BW) (d) diabetic rat which received PWCG at dose of 100mgsdotkgminus1 BW(DM + PWCG 100mgsdotkgminus1 BW) (e) diabetic rat which received PWCG at dose of 200mgsdotkgminus1 BW (DM + PWCG 200mgsdotkgminus1 BW)

the elevation of both CAT and GPx might be responsible forthe decreased oxidative stress which in turn enhanced thesurvival of ganglion cells in OUL leading to the increasedtotal retina thickness However the decreased oxidative stressin rat lens induced by medium dose of PWCG might beassociated with other mechanism in addition to the elevationof the scavenger enzymes activities Since the excess oxidativestress occurs as the result of imbalance between oxidativestress formation and oxidative stress buffering capacities wesuggested that the decreased oxidative stress formationmightplay a role on the reduction of oxidative stress induced by thePWCG especially at the medium concentration

Oxidative stress and aldose reductase also play the crucialrole on the cataractogenesis in diabetic rats [24 25] In addi-tion it has been reported that antioxidant and aldose reduc-tase inhibitory effects can improve cataract [26] Thus theanticataractogenis of PWCG might also be associated withboth effects mentioned earlier Aldose reductase also playsa role on the pathophysiology of diabetic retinopathy [27]

Based on the basis of this information we do suggest thatthe effect of PWCG to improve retinopathy especially at highdose should also relate with the aldose reductase inhibitoryeffect

Previous study has demonstrated that flavonoids canprotect against oxidative stress and increase the survivalof ganglion cells [28] and prevent cataract [29] in diabeticrats Recent study has demonstrated that anthocyanins andanthocyanins-rich substance have the potential to protectagainst retinopathy [30] and diabetic cataract [26] In addi-tion ginger and its constituents also show the potential toprotect against diabetic retinopathy [31] Therefore the anti-cataractogenesis and antiretinopathy of PWCGmay be due tothe flavonoids such as anthocyanins in PWCG Other typesof flavonoidmay also play a role However further researchesare required to confirm the possible active ingredient

In this study the slit lamp examination clearly providedthe changes of lens characteristic However no pupil wasdilated during the grading of severity as that was performed

8 Oxidative Medicine and Cellular Longevity

0002000400060008000

100001200014000160001800020000

lowastlowastlowastlowastlowastlowast

lowastlowastlowast

Tota

l ret

inal

thic

knes

s (120583

m)

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 6 The total retinal thickness of retina (TRT) of control anddiabetic rats (DM) which received either vehicle or the combinationextract of purple waxy corn and ginger (PWCG) at doses of 50 100and 200mgsdotkgminus1 BW for 10weeks 119875 valuelt 0001 comparedwithcontrol and lowastlowastlowast119875 value lt 0001 compared with vehicle

000500

100015002000250030003500400045005000

The r

etin

al n

ucle

ar la

yer t

hick

ness

(120583m

)

lowastlowastlowast lowastlowastlowastlowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 7 The thickness of the retinal outer nuclear layer (ROT)of control and diabetic rats (DM) which received either vehicle orthe combination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks

119875 valuelt 0001 compared with control and lowastlowastlowast119875 value lt 0001 comparedwith vehicle

during the slit lamp examination in humanThismay possiblyinfluence the precision of grading scores which reflect thereal severity of cataract To obtain the precise severity ofcataract the slit lamp examination should be performedwhile pupil was dilated However all rats were assessed underthe condition without the pupil dilator so no confoundingerror about the beneficial effect of PWCG on cataract wasachieved

The lenticular opacification in control rats was alsoobserved during the 6ndash10 weeks of study period Based onthe previous study that stress hormone or glucocorticoids

000500

100015002000250030003500400045005000

The n

umbe

r of n

euro

nin

gan

glio

n ce

ll lay

er in

retin

a

lowastlowastlowastlowastlowastlowast

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 8 The number of cells in the ganglion cell layer (NG)in retina of control and diabetic rats (DM) which received eithervehicle or the combination extract of purple waxy corn and ginger(PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks 119875value lt 0001 compared with control and lowast lowastlowastlowast119875 value lt 005 and0001 compared with vehicle

MD

A (U

mg

prot

ein)

000

100

200

300

400

500

600

lowastlowastlowast lowastlowastlowastlowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 9 The level of malondialdehyde (MDA) (Umg protein) inlens of control and diabetic rats (DM) which received either vehicleor the combination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 = 8group)119875 value lt 0001 compared with control and lowastlowastlowast119875 value lt 0001

compared with diabetic rats which received vehicle (DM + vehicle)

can also induce cataract [32 33] we do suggest that thedevelopment of cataract in control group may possibly occuras the exposure to stress condition leading to the elevationof glucocorticoids hormone resulting in the cataractogenesisUnfortunately the level of corticosterone has not been deter-mined in this study Therefore the precise understandingmechanism of this change requires further investigation Inaddition spontaneous age-related cataract can also occurin laboratory rodents The prevalence is varied dependingon species Even young adolescent rodents can be found tohave cataract approximately 20 [34]Therefore the cataract

Oxidative Medicine and Cellular Longevity 9SO

D (U

mg

prot

ein)

000050100150200250300350400450

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 10 The activity of superoxide dismutase (SOD) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BWfor 10weeks(119873 = 8group) 119875 value lt 0001 compared with control

CAT

(Um

g pr

otei

n)

000

1000

2000

3000

4000

5000

6000

lowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 11 The activity of catalase (CAT) (Umg protein) in lensof control and diabetic rats (DM) which received either vehicle orthe combination extract of purple waxy corn and ginger (PWCG) atdoses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 = 8group)119875 value lt 0001 compared with control and lowastlowast119875 value lt 001

compared with diabetic rats which received vehicle (DM + vehicle)

observed in normal rats might occur either via stress orvia spontaneous cataractogenesis of the laboratory rodentsmentioned earlier

5 Conclusion

The combination extract of purple waxy corn seeds andginger rhizome (PWCG) can protect against diabetic cataractand diabetic retinopathyThepossible underlyingmechanismmay occur partly via the decreased oxidative stress and thesuppression of aldose reductase activity in lens The possibleactive ingredient may be associated with flavonoids such as

000020040060080100120140160180200

GPx

(Um

g pr

otei

n)

lowastlowastlowast

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 12 The activity of glutathione peroxidase (GPx) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10weeks (119873 = 8group) 119875 value lt 0001 compared with controland lowast lowastlowastlowast119875 value lt 005 0001 respectively compared with diabeticrats which received vehicle (DM + vehicle)

AR

activ

ity (n

mol

min

mg)

00000

00010

00020

00030

00040

00050

00060

00070

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 13 The activity of aldose reductase (AR) (nmolminmg)in lens of control and diabetic rats (DM) which received eithervehicle or the combination extract of purple waxy corn and ginger(PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 =8group) 119875 value lt 005 compared with control and lowast119875 value lt005 compared with diabetic rats which received vehicle (DM +vehicle)

anthocyanin and other flavonoids Further researches are stillessential to understand the precise underlying mechanismand possible active ingredient

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

10 Oxidative Medicine and Cellular Longevity

Acknowledgments

This study was supported by the Higher Education ResearchPromotion andNational Research University Project ofThai-land Office of the Higher Education Commission throughthe Food and Functional Food Research Cluster of KhonKaen University and the Integrative Complementary Alter-native Medicine Research and Development Center KhonKaen University Thailand Great thanks were also extendedto Research and Development Center of Herbal HealthProduct for providing the opportunity to collaborate in thesubcluster

References

[1] R L Engerman and T S Kern ldquoHyperglycemia as a cause ofdiabetic retinopathyrdquo Metabolism vol 35 supplement 1 no 4pp 20ndash23 1986

[2] A Pollreisz and U Schmidt-Erfurth ldquoDiabetic cataractmdashpathogenesis epidemiology and treatmentrdquo Journal of Ophthal-mology vol 2010 Article ID 608751 8 pages 2010

[3] I G Obrosova S S M Chung and P F Kador ldquoDiabeticcataracts mechanisms and managementrdquo DiabetesMetabolismResearch and Reviews vol 26 no 3 pp 172ndash180 2010

[4] J Ding and T Y Wong ldquoCurrent epidemiology of diabeticretinopathy and diabetic macular edemardquo Current DiabetesReports vol 12 no 4 pp 346ndash354 2012

[5] A Y W Lee and S S M Chung ldquoContributions of polyolpathway to oxidative stress in diabetic cataractrdquo The FASEBJournal vol 13 no 1 pp 23ndash30 1999

[6] S A Madsen-Bouterse and R A Kowluru ldquoOxidative stressand diabetic retinopathy pathophysiological mechanisms andtreatment perspectivesrdquo Reviews in Endocrine and MetabolicDisorders vol 9 no 4 pp 315ndash327 2008

[7] I G Obrosova and P F Kador ldquoAldose reductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 1389ndash2010 2011

[8] V R Drel P Pacher T K Ali et al ldquoAldose reductase inhibitorfidarestat counteracts diabetes-associated cataract formationretinal oxidative-nitrosative stress glial activation and apopto-sisrdquo International Journal of Molecular Medicine vol 21 no 6pp 667ndash676 2008

[9] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (Purple Waxy Corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[10] Y Morimitsu K Kubota T Tashiro E Hashizume T Kamiyaand T Osawa ldquoInhibitory effect of anthocyanins and coloredrice ondiabetic cataract formation in the rat lensesrdquo Interna-tional Congress Series vol 1245 pp 503ndash508 2002

[11] M Saraswat P Suryanarayana P Y Reddy M A Patil NBalakrishna and G B Reddy ldquoAntiglycating potential of Zin-giber officinalis and delay of diabetic cataract in ratsrdquoMolecularVision vol 16 pp 1525ndash1537 2010

[12] Y Li V H Tran C C Duke and B D Roufogalis ldquoPreventiveand protective properties of Zingiber officinale (Ginger) indiabetes mellitus diabetic complications and associated lipidand other metabolic disorders a brief reviewrdquo Evidence-basedComplementary and Alternative Medicine vol 2012 Article ID516870 10 pages 2012

[13] P Suryanarayana M Saraswat T Mrudula T P Krishna KKrishnaswamy andG B Reddy ldquoCurcumin and turmeric delaystreptozotocin-induced diabetic cataract in ratsrdquo InvestigativeOphthalmology amp Visual Science vol 46 no 6 pp 2092ndash20992005

[14] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[15] Y Sun L W Oberley and Y Li ldquoA simple method for clinicalassay of superoxide dismutaserdquo Clinical Chemistry vol 34 no3 pp 497ndash500 1988

[16] L Goth ldquoA simple method for determination of serum catalaseactivity and revision of reference rangerdquo Clinica Chimica Actavol 196 no 2-3 pp 143ndash151 1991

[17] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973

[18] M B Patel and S M Mishra ldquoAldose reductase inhibitoryactivity and anti catraract potential of some traditionallyacclaimed antidiabetic medicinal plantsrdquo Oriental Pharmacyand Experimental Medicine vol 9 no 3 pp 245ndash251 2009

[19] R Agarwal I Iezhitsa N A Awaludin et al ldquoEffects ofmagnesium taurate on the onset and progression of galactose-induced experimental cataract in vivo and in vitro evaluationrdquoExperimental Eye Research vol 110 pp 35ndash43 2013

[20] P M Martin P Roon T K van Ells V Ganapathy and SB Smith ldquoDeath of retinal neurons in streptozotocin-induceddiabetic micerdquo Investigative Ophthalmology and Visual Sciencevol 45 no 9 pp 3330ndash3336 2004

[21] H W van Dijk F D Verbraak P H B Kok et al ldquoDecreasedretinal ganglion cell layer thickness in patients with type 1diabetesrdquo Investigative Ophthalmology and Visual Science vol51 no 7 pp 3660ndash3665 2010

[22] T S Kern and A J Barber ldquoRetinal ganglion cells in diabetesrdquoJournal of Physiology vol 586 no 18 pp 4401ndash4408 2008

[23] G A Ramadan ldquoSorbitol-induced diabetic-like retinal lesionsin rats microscopic studyrdquo American Journal of Pharmacologyand Toxicology vol 2 no 2 pp 89ndash97 2007

[24] B Ozmen D Ozmen E Erkin I Guner S Habif and OBaymdir ldquoLens superoxide dismutase and catalase activities indiabetic cataractrdquoClinical Biochemistry vol 35 no 1 pp 69ndash722002

[25] A B M Reddy R Tammali R Mishra S Srivastava S KSrivastava and K V Ramana ldquoAldose reductase deficiencyprotects sugar-induced lens opacification in ratsrdquo Chemico-Biological Interactions vol 191 no 1ndash3 pp 346ndash350 2011

[26] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (purple waxy corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[27] I G Obrosova and P F Kador ldquoAldosereductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 373ndash385 2011

[28] PMaher andAHanneken ldquoFlavonoids protect retinal ganglioncells from oxidative stress-induced deathrdquo Investigative Oph-thalmology and Visual Science vol 46 no 12 pp 4796ndash48032005

[29] M Stefek ldquoNatural flavonoids as potential multifunctionalagents in prevention of diabetic cataractrdquo InterdisciplinaryToxicology vol 4 no 2 pp 69ndash77 2011

Oxidative Medicine and Cellular Longevity 11

[30] S F Nabavi S Habtemariam M Daglia N Shafighi A JBarber and S M Nabavi ldquoAnthocyanins as a potential therapyfor diabetic retinopathyrdquo Current Medicinal Chemistry vol 22no 1 pp 51ndash58 2014

[31] A H Rahmani F M Al Shabrmi and S M Aly ldquoActiveingredients of ginger as potential candidates in the preventionand treatment of diseases via modulation of biological activ-itiesrdquo International Journal of Physiology Pathophysiology andPharmacology vol 6 no 2 pp 125ndash136 2014

[32] H Watanabe H Kosano and H Nishigori ldquoSteroid-inducedshort term diabetes in chick embryo reversible effects of insulinon metabolic changes and cataract formationrdquo InvestigativeOphthalmology amp Visual Science vol 41 no 7 pp 1846ndash18522000

[33] E R James ldquoThe etiology of steroid cataractrdquo Journal of OcularPharmacology and Therapeutics vol 23 no 5 pp 403ndash4202007

[34] P Greaves Histopathological of Preclinical Toxicity StudiesElsevier New York NY USA 3rd edition 2007

Submit your manuscripts athttpwwwhindawicom

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Behavioural Neurology

EndocrinologyInternational Journal of

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Disease Markers

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OncologyJournal of

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

4 Oxidative Medicine and Cellular Longevity

Table 1 Opacity index of the lens of normal and diabetic rats (DM) which received either vehicle or PWCG at doses of 50 100 and200mgsdotkgminus1 BW for 10 weeks (119873 = 8group) 119875 value lt 001 and 0001 respectively compared with control group lowast lowastlowast lowastlowastlowast119875 valuelt 005 001 and 0001 respectively compared with diabetic rats which received vehicle (DM + vehicle)

Time durationOpacity index of the lens (mean plusmn SEM)

Control DM + vehicle DM + PWCG(50mgkgsdotBWminus1)

DM + PWCG(100mgkgsdotBWminus1)

DM + PWCG(200mgkgsdotBWminus1)

Baseline 000 plusmn 000 000 plusmn 000 000 plusmn 000 000 plusmn 000 000 plusmn 000Week 1 000 plusmn 000 000 plusmn 000 000 plusmn 000 000 plusmn 000 000 plusmn 000Week 2 000 plusmn 000 006 plusmn 006 000 plusmn 000 000 plusmn 000 000 plusmn 000Week 3 000 plusmn 000 050 plusmn 016 014 plusmn 010lowast 050 plusmn 013 022 plusmn 010Week 4 000 plusmn 000 100 plusmn 020 079 plusmn 024 100 plusmn 018 072 plusmn 023Week 5 000 plusmn 000 181 plusmn 031 100 plusmn 021lowast 125 plusmn 023 100 plusmn 028lowast

Week 6 008 plusmn 008 225 plusmn 032 150 plusmn 029 150 plusmn 022 111 plusmn 031lowastlowast

Week 7 017 plusmn 011 250 plusmn 034 171 plusmn 029 188 plusmn 029 144 plusmn 041lowast

Week 8 025 plusmn 013 306 plusmn 023 200 plusmn 030lowast 219 plusmn 025lowast 167 plusmn 040lowastlowastlowast

Week 9 050 plusmn 015 381 plusmn 010 229 plusmn 035lowastlowastlowast 238 plusmn 026lowastlowastlowast 178 plusmn 039lowastlowastlowast

Week 10 050 plusmn 015 400 plusmn 000 221 plusmn 037lowastlowastlowast 263 plusmn 026lowastlowastlowast 189 plusmn 040lowastlowastlowast

000

10000

20000

30000

40000

50000

60000

Bloo

d gl

ucos

e lev

el (m

gD

L)

ControlDM + vehicle

Time duration

Week 1 Week 5 Week 10

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 1 The average fasting blood glucose levels of all the experi-mental groups at 1-week 5-week and 10-week intervention period(119873 = 8group) 119875 value lt 0001 compared with control group

period PWCGat all doses used in this study failed to decreasethe fasting blood glucose level in diabetic rats

32 Anticataract Effect of PWCG Theeffect of PWCGon lensopacity was shown in Figure 2The lens opacity in control ratsalso developed cataract during 6ndash10-week period At 10-weekstudy period all lens from both right (1A and 1B) and left eyes(1a and 1b) of control rats were clear and normal Both right(2A and 2B) and left eyes (2a and 2b) of diabetic rats whichreceived vehicle showed lens opacity However the increasedlens opacity induced by diabetic condition was attenuated indiabetic rats which received all doses of PWCG (3Andash5A 3Bndash5B 3andash5a and 3bndash5b)

Table 1 showed that the lens opacity indices of diabeticrats which received vehicle started to increase at 3-week

treatment duration (119875 value lt 001 compared with controlrats) This change was still observed until the end of study(119875 value lt 0001 compared with control rats) Diabetic ratswhich received PWCG at dose of 200mgsdotkgminus1 significantlymitigated the elevation of lens opacity index induced by dia-betes at 5-week treatment (119875 valuelt 005 comparedwith dia-betic rats which received vehicle)This change persisted untilthe end of study (119875 value lt 001 005 0001 0001 0001 respcomparedwith diabetic rats which received vehicle) Diabeticrats which received PWCG at low dose (50mgsdotkgminus1 BW)also mitigated the elevation of lens opacity index induced bydiabetes but the significant changes were observed at 3-week5 week 8-week 9-week and 10-week treatment duration (119875value lt 005 005 005 0001 and 0001 resp compared withdiabetic rats which received vehicle) However the mediumdose of PWCG produced the significant mitigation effect onlens opacity index induced by diabetes only at 8-week 9-week and 10-week treatment duration (119875 value lt 005 0001and 0001 resp compared with diabetic rats which receivedvehicle)The effects of PWCGon lens opacity were confirmedby histopathological analysis of lens as shown in Figure 3It was found that at 10-week study period the lens capsulethickness and the density of epithelial cells of diabetic ratsubjected to vehicle treatment were decreased

In addition the reduction of differentiated lens fibersvacuoles and homogenized area was also observed (Figure3(b)) Diabetic rats which received PWCG at doses of 50100 and 200mgsdotkgminus1 BW showed the increased density ofepithelial cells and differentiated lens fiber thickness togetherwith the decreased vacuoles and homogenized area (Figures3(c) 3(d) and 3(e))

Figure 4 showed the effect of PWCG on the severity oflens damage Our results showed that the diabetic rats whichreceived vehicle showed the increased severity of cataract(119875 value lt 0001 compared with control rats) All doses ofPWCG significantly mitigated the enhanced cataract severityinduced by diabetes mellitus (119875 value lt 0001 all comparedwith diabetic rats which received vehicle)

Oxidative Medicine and Cellular Longevity 5

(1A) (2A) (3A) (4A) (5A)

(1B) (2B) (3B) (4B) (5B)

(1a) (2a) (3a) (4a) (5a)

(1b) (2b) (3b) (4b) (5b)

Figure 2 Representative photographs showing the effects of 10-week treatment of PWCG in prevention of cataract A photographs via slitlamp B photographs via camera 1 control rat 2 diabetic rat which received vehicle (DM + vehicle) 3 diabetic rat which received PWCGat dose of 50mgsdotkgminus1 BW (DM + PWCG 50mgsdotkgminus1 BW) 4 diabetic rat which received PWCG at dose of 100mgsdotkgminus1 BW (DM + PWCG100mgsdotkgminus1 BW) 5 diabetic rat which received PWCG at dose of 200mgsdotkgminus1 BW (DM + PWCG 200mgsdotkgminus1 BW)

33 The Effect of PWCG on Retinopathy in Diabetic Rats Theeffect of PWCG on histopathological change of retina wasshown in Figure 5 At 10-week study period the total retinalthickness (TRT) the thickness of the retinal outer nuclearlayer (ROT) and the number of cells in the ganglion celllayer (NG) of diabetic ratswhich received vehicle significantlydecreasedwhen compared to controlHowever the decreasedTRT ROT and NG induced by diabetic condition wereattenuated in diabetic rats which received all doses of PWCGFigures 6 7 and 8 showed that diabetic rats which receivedPWCG at doses of 50 100 and 200mgsdotkgminus1 BW significantlyincreased the TRT (119875 value lt 0001 all compared withdiabetic rats which received vehicle) ROT (119875 value lt 0001all compared with diabetic rats which received vehicle) andNG of retina (119875 value lt 0001 005 and 0001 resp comparedwith diabetic rats which received vehicle)

34 Effect of PWCG on Oxidative Stress Markers Based onthe crucial role of oxidative stress on cataract and retinopathythe effects of PWCG on oxidative stress markers includinglevel of MDA and the activities of SOD CAT and GPx inlens were carried out The results were shown in Figures 9

10 11 and 12 It was found that diabetic rats which receivedvehicle significantly increased MDA level (119875 value lt 0001compared with control) but decreased SOD CAT and GPxactivities (119875 value lt 0001 all compared with control) in lensPWCG at doses of 50 100 and 200mgsdotkgminus1 BW significantlymitigated the elevation of MDA level induced by diabetesmellitus (119875 value lt 0001 all compared with diabetic ratswhich received vehicle) Low dose of PWCG also mitigatedthe reduction of GPx activity in lens of diabetic rats (119875value lt 005 compared with diabetic rats which receivedvehicle) whereas high dose of PWCGmitigated the reductionof both CAT and GPx activities of lens of diabetic rats (119875value lt 001 and 0001 resp compared with diabetic ratswhich received vehicle) However no significant changes ofscavenger enzymes activities were observed in the diabeticrats which received medium dose of PWCG

35 Effect of PWCG on Aldose Reductase Activity In additionto the oxidative stress the polyol pathway also plays thecrucial role on the cataractogenesis Therefore the effectof PWCG on aldose reductase enzyme activity in lenswas also investigated and data were shown in Figure 13 It

6 Oxidative Medicine and Cellular Longevity

50 120583m

(a)

50 120583m

(b)

50 120583m

(c)

50 120583m

(d)

50 120583m

(e)

Figure 3 Photographs of transverse sections of eye balls (5 120583m thickness) stained with HampE which determined the cataract severity at theend of experiment using light microscope (week 10) (a) Control (b) DM + Vehicle (c) DM + PWCG (50mgkgsdotBW) (d) DM + PWCG(100mgkgsdotBW) and (e) DM + PWCG (200mgkgsdotBW)

000

050

100

150

200

250

300

350

400

Gra

de o

f cat

arac

t sev

erity

lowastlowastlowast

lowastlowastlowast

lowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 4 The cataract severity of control and diabetic rats (DM)which received either vehicle or PWCG at doses of 50 100 and200mgsdotkgminus1 BW for 10 weeks

119875 value lt 0001 compared withcontrol and lowastlowastlowast119875 value lt 0001 compared with vehicle

was demonstrated that aldose reductase activity in lens ofdiabetic rats was increased (119875 value lt 005 compared withcontrol) PWCG at dose of 200mgsdotkgminus1 BW could signifi-cantly decrease the elevation of aldose reductase induced bydiabetes mellitus in rat lens No other significant changeswere observed

4 Discussion

Diabetic cataract and retinopathy are the important causes ofblindness so the prevention of these conditions is very muchimportant Our results clearly demonstrated that PWCG sig-nificantly mitigated the reduction of ganglia in outer nuclearlayer (ONL) OUL thickness and total retina thickness Inaddition PWCG also showed anticataract effect Togetherwith the changes mentioned earlier the decreased oxidativestress and aldose reductase activity in rat lens were alsoobserved

It has been reported that the neurodegeneration of gan-glion cells plays the crucial role on the reduction of OULand total retina thickness [21] The degeneration of retinalganglion cells is under the influence of many factors suchas inflammation oxidative stress or exposure to advancedglycation end products [22] and polyol pathway [23] Ourdata also demonstrates the increased oxidative stress markerssuch asMDA level and the enhanced aldose reductase activityin lens of diabetic rats These changes are in agreement withthe previous study [22 23] Therefore we suggested thatthe effect of PWCG to attenuate retinopathy induced bydiabetes mellitus might be partly related to the decreasedoxidative stress and aldose reductase activity in rat lens Itwas found that the enhanced of GPx activity in rat lens mightbe responsible in part for the reduction of oxidative stress inrat lens of diabetic rats especially at low and high doses ofPWCG In addition to the increased GPx activity the highdose of PWCG also enhanced CAT in rat lens Therefore

Oxidative Medicine and Cellular Longevity 7

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(a)

50 120583m

RODS and CONES

ONL

OPLINL

IPLGCL

(b)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(c)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(d)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(e)

Figure 5 Light microscope of transverse section of eye ball (5 120583m thickness) stained with HampE which determined the pathological changeof retina at the end of experiment (week 10) (a) Control rat (b) diabetic rat which received vehicle (DM + vehicle) (c) diabetic rat whichreceived PWCG at dose of 50mgsdotkgminus1 BW (DM + PWCG 50mgsdotkgminus1 BW) (d) diabetic rat which received PWCG at dose of 100mgsdotkgminus1 BW(DM + PWCG 100mgsdotkgminus1 BW) (e) diabetic rat which received PWCG at dose of 200mgsdotkgminus1 BW (DM + PWCG 200mgsdotkgminus1 BW)

the elevation of both CAT and GPx might be responsible forthe decreased oxidative stress which in turn enhanced thesurvival of ganglion cells in OUL leading to the increasedtotal retina thickness However the decreased oxidative stressin rat lens induced by medium dose of PWCG might beassociated with other mechanism in addition to the elevationof the scavenger enzymes activities Since the excess oxidativestress occurs as the result of imbalance between oxidativestress formation and oxidative stress buffering capacities wesuggested that the decreased oxidative stress formationmightplay a role on the reduction of oxidative stress induced by thePWCG especially at the medium concentration

Oxidative stress and aldose reductase also play the crucialrole on the cataractogenesis in diabetic rats [24 25] In addi-tion it has been reported that antioxidant and aldose reduc-tase inhibitory effects can improve cataract [26] Thus theanticataractogenis of PWCG might also be associated withboth effects mentioned earlier Aldose reductase also playsa role on the pathophysiology of diabetic retinopathy [27]

Based on the basis of this information we do suggest thatthe effect of PWCG to improve retinopathy especially at highdose should also relate with the aldose reductase inhibitoryeffect

Previous study has demonstrated that flavonoids canprotect against oxidative stress and increase the survivalof ganglion cells [28] and prevent cataract [29] in diabeticrats Recent study has demonstrated that anthocyanins andanthocyanins-rich substance have the potential to protectagainst retinopathy [30] and diabetic cataract [26] In addi-tion ginger and its constituents also show the potential toprotect against diabetic retinopathy [31] Therefore the anti-cataractogenesis and antiretinopathy of PWCGmay be due tothe flavonoids such as anthocyanins in PWCG Other typesof flavonoidmay also play a role However further researchesare required to confirm the possible active ingredient

In this study the slit lamp examination clearly providedthe changes of lens characteristic However no pupil wasdilated during the grading of severity as that was performed

8 Oxidative Medicine and Cellular Longevity

0002000400060008000

100001200014000160001800020000

lowastlowastlowastlowastlowastlowast

lowastlowastlowast

Tota

l ret

inal

thic

knes

s (120583

m)

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 6 The total retinal thickness of retina (TRT) of control anddiabetic rats (DM) which received either vehicle or the combinationextract of purple waxy corn and ginger (PWCG) at doses of 50 100and 200mgsdotkgminus1 BW for 10weeks 119875 valuelt 0001 comparedwithcontrol and lowastlowastlowast119875 value lt 0001 compared with vehicle

000500

100015002000250030003500400045005000

The r

etin

al n

ucle

ar la

yer t

hick

ness

(120583m

)

lowastlowastlowast lowastlowastlowastlowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 7 The thickness of the retinal outer nuclear layer (ROT)of control and diabetic rats (DM) which received either vehicle orthe combination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks

119875 valuelt 0001 compared with control and lowastlowastlowast119875 value lt 0001 comparedwith vehicle

during the slit lamp examination in humanThismay possiblyinfluence the precision of grading scores which reflect thereal severity of cataract To obtain the precise severity ofcataract the slit lamp examination should be performedwhile pupil was dilated However all rats were assessed underthe condition without the pupil dilator so no confoundingerror about the beneficial effect of PWCG on cataract wasachieved

The lenticular opacification in control rats was alsoobserved during the 6ndash10 weeks of study period Based onthe previous study that stress hormone or glucocorticoids

000500

100015002000250030003500400045005000

The n

umbe

r of n

euro

nin

gan

glio

n ce

ll lay

er in

retin

a

lowastlowastlowastlowastlowastlowast

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 8 The number of cells in the ganglion cell layer (NG)in retina of control and diabetic rats (DM) which received eithervehicle or the combination extract of purple waxy corn and ginger(PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks 119875value lt 0001 compared with control and lowast lowastlowastlowast119875 value lt 005 and0001 compared with vehicle

MD

A (U

mg

prot

ein)

000

100

200

300

400

500

600

lowastlowastlowast lowastlowastlowastlowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 9 The level of malondialdehyde (MDA) (Umg protein) inlens of control and diabetic rats (DM) which received either vehicleor the combination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 = 8group)119875 value lt 0001 compared with control and lowastlowastlowast119875 value lt 0001

compared with diabetic rats which received vehicle (DM + vehicle)

can also induce cataract [32 33] we do suggest that thedevelopment of cataract in control group may possibly occuras the exposure to stress condition leading to the elevationof glucocorticoids hormone resulting in the cataractogenesisUnfortunately the level of corticosterone has not been deter-mined in this study Therefore the precise understandingmechanism of this change requires further investigation Inaddition spontaneous age-related cataract can also occurin laboratory rodents The prevalence is varied dependingon species Even young adolescent rodents can be found tohave cataract approximately 20 [34]Therefore the cataract

Oxidative Medicine and Cellular Longevity 9SO

D (U

mg

prot

ein)

000050100150200250300350400450

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 10 The activity of superoxide dismutase (SOD) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BWfor 10weeks(119873 = 8group) 119875 value lt 0001 compared with control

CAT

(Um

g pr

otei

n)

000

1000

2000

3000

4000

5000

6000

lowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 11 The activity of catalase (CAT) (Umg protein) in lensof control and diabetic rats (DM) which received either vehicle orthe combination extract of purple waxy corn and ginger (PWCG) atdoses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 = 8group)119875 value lt 0001 compared with control and lowastlowast119875 value lt 001

compared with diabetic rats which received vehicle (DM + vehicle)

observed in normal rats might occur either via stress orvia spontaneous cataractogenesis of the laboratory rodentsmentioned earlier

5 Conclusion

The combination extract of purple waxy corn seeds andginger rhizome (PWCG) can protect against diabetic cataractand diabetic retinopathyThepossible underlyingmechanismmay occur partly via the decreased oxidative stress and thesuppression of aldose reductase activity in lens The possibleactive ingredient may be associated with flavonoids such as

000020040060080100120140160180200

GPx

(Um

g pr

otei

n)

lowastlowastlowast

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 12 The activity of glutathione peroxidase (GPx) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10weeks (119873 = 8group) 119875 value lt 0001 compared with controland lowast lowastlowastlowast119875 value lt 005 0001 respectively compared with diabeticrats which received vehicle (DM + vehicle)

AR

activ

ity (n

mol

min

mg)

00000

00010

00020

00030

00040

00050

00060

00070

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 13 The activity of aldose reductase (AR) (nmolminmg)in lens of control and diabetic rats (DM) which received eithervehicle or the combination extract of purple waxy corn and ginger(PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 =8group) 119875 value lt 005 compared with control and lowast119875 value lt005 compared with diabetic rats which received vehicle (DM +vehicle)

anthocyanin and other flavonoids Further researches are stillessential to understand the precise underlying mechanismand possible active ingredient

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

10 Oxidative Medicine and Cellular Longevity

Acknowledgments

This study was supported by the Higher Education ResearchPromotion andNational Research University Project ofThai-land Office of the Higher Education Commission throughthe Food and Functional Food Research Cluster of KhonKaen University and the Integrative Complementary Alter-native Medicine Research and Development Center KhonKaen University Thailand Great thanks were also extendedto Research and Development Center of Herbal HealthProduct for providing the opportunity to collaborate in thesubcluster

References

[1] R L Engerman and T S Kern ldquoHyperglycemia as a cause ofdiabetic retinopathyrdquo Metabolism vol 35 supplement 1 no 4pp 20ndash23 1986

[2] A Pollreisz and U Schmidt-Erfurth ldquoDiabetic cataractmdashpathogenesis epidemiology and treatmentrdquo Journal of Ophthal-mology vol 2010 Article ID 608751 8 pages 2010

[3] I G Obrosova S S M Chung and P F Kador ldquoDiabeticcataracts mechanisms and managementrdquo DiabetesMetabolismResearch and Reviews vol 26 no 3 pp 172ndash180 2010

[4] J Ding and T Y Wong ldquoCurrent epidemiology of diabeticretinopathy and diabetic macular edemardquo Current DiabetesReports vol 12 no 4 pp 346ndash354 2012

[5] A Y W Lee and S S M Chung ldquoContributions of polyolpathway to oxidative stress in diabetic cataractrdquo The FASEBJournal vol 13 no 1 pp 23ndash30 1999

[6] S A Madsen-Bouterse and R A Kowluru ldquoOxidative stressand diabetic retinopathy pathophysiological mechanisms andtreatment perspectivesrdquo Reviews in Endocrine and MetabolicDisorders vol 9 no 4 pp 315ndash327 2008

[7] I G Obrosova and P F Kador ldquoAldose reductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 1389ndash2010 2011

[8] V R Drel P Pacher T K Ali et al ldquoAldose reductase inhibitorfidarestat counteracts diabetes-associated cataract formationretinal oxidative-nitrosative stress glial activation and apopto-sisrdquo International Journal of Molecular Medicine vol 21 no 6pp 667ndash676 2008

[9] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (Purple Waxy Corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[10] Y Morimitsu K Kubota T Tashiro E Hashizume T Kamiyaand T Osawa ldquoInhibitory effect of anthocyanins and coloredrice ondiabetic cataract formation in the rat lensesrdquo Interna-tional Congress Series vol 1245 pp 503ndash508 2002

[11] M Saraswat P Suryanarayana P Y Reddy M A Patil NBalakrishna and G B Reddy ldquoAntiglycating potential of Zin-giber officinalis and delay of diabetic cataract in ratsrdquoMolecularVision vol 16 pp 1525ndash1537 2010

[12] Y Li V H Tran C C Duke and B D Roufogalis ldquoPreventiveand protective properties of Zingiber officinale (Ginger) indiabetes mellitus diabetic complications and associated lipidand other metabolic disorders a brief reviewrdquo Evidence-basedComplementary and Alternative Medicine vol 2012 Article ID516870 10 pages 2012

[13] P Suryanarayana M Saraswat T Mrudula T P Krishna KKrishnaswamy andG B Reddy ldquoCurcumin and turmeric delaystreptozotocin-induced diabetic cataract in ratsrdquo InvestigativeOphthalmology amp Visual Science vol 46 no 6 pp 2092ndash20992005

[14] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[15] Y Sun L W Oberley and Y Li ldquoA simple method for clinicalassay of superoxide dismutaserdquo Clinical Chemistry vol 34 no3 pp 497ndash500 1988

[16] L Goth ldquoA simple method for determination of serum catalaseactivity and revision of reference rangerdquo Clinica Chimica Actavol 196 no 2-3 pp 143ndash151 1991

[17] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973

[18] M B Patel and S M Mishra ldquoAldose reductase inhibitoryactivity and anti catraract potential of some traditionallyacclaimed antidiabetic medicinal plantsrdquo Oriental Pharmacyand Experimental Medicine vol 9 no 3 pp 245ndash251 2009

[19] R Agarwal I Iezhitsa N A Awaludin et al ldquoEffects ofmagnesium taurate on the onset and progression of galactose-induced experimental cataract in vivo and in vitro evaluationrdquoExperimental Eye Research vol 110 pp 35ndash43 2013

[20] P M Martin P Roon T K van Ells V Ganapathy and SB Smith ldquoDeath of retinal neurons in streptozotocin-induceddiabetic micerdquo Investigative Ophthalmology and Visual Sciencevol 45 no 9 pp 3330ndash3336 2004

[21] H W van Dijk F D Verbraak P H B Kok et al ldquoDecreasedretinal ganglion cell layer thickness in patients with type 1diabetesrdquo Investigative Ophthalmology and Visual Science vol51 no 7 pp 3660ndash3665 2010

[22] T S Kern and A J Barber ldquoRetinal ganglion cells in diabetesrdquoJournal of Physiology vol 586 no 18 pp 4401ndash4408 2008

[23] G A Ramadan ldquoSorbitol-induced diabetic-like retinal lesionsin rats microscopic studyrdquo American Journal of Pharmacologyand Toxicology vol 2 no 2 pp 89ndash97 2007

[24] B Ozmen D Ozmen E Erkin I Guner S Habif and OBaymdir ldquoLens superoxide dismutase and catalase activities indiabetic cataractrdquoClinical Biochemistry vol 35 no 1 pp 69ndash722002

[25] A B M Reddy R Tammali R Mishra S Srivastava S KSrivastava and K V Ramana ldquoAldose reductase deficiencyprotects sugar-induced lens opacification in ratsrdquo Chemico-Biological Interactions vol 191 no 1ndash3 pp 346ndash350 2011

[26] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (purple waxy corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[27] I G Obrosova and P F Kador ldquoAldosereductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 373ndash385 2011

[28] PMaher andAHanneken ldquoFlavonoids protect retinal ganglioncells from oxidative stress-induced deathrdquo Investigative Oph-thalmology and Visual Science vol 46 no 12 pp 4796ndash48032005

[29] M Stefek ldquoNatural flavonoids as potential multifunctionalagents in prevention of diabetic cataractrdquo InterdisciplinaryToxicology vol 4 no 2 pp 69ndash77 2011

Oxidative Medicine and Cellular Longevity 11

[30] S F Nabavi S Habtemariam M Daglia N Shafighi A JBarber and S M Nabavi ldquoAnthocyanins as a potential therapyfor diabetic retinopathyrdquo Current Medicinal Chemistry vol 22no 1 pp 51ndash58 2014

[31] A H Rahmani F M Al Shabrmi and S M Aly ldquoActiveingredients of ginger as potential candidates in the preventionand treatment of diseases via modulation of biological activ-itiesrdquo International Journal of Physiology Pathophysiology andPharmacology vol 6 no 2 pp 125ndash136 2014

[32] H Watanabe H Kosano and H Nishigori ldquoSteroid-inducedshort term diabetes in chick embryo reversible effects of insulinon metabolic changes and cataract formationrdquo InvestigativeOphthalmology amp Visual Science vol 41 no 7 pp 1846ndash18522000

[33] E R James ldquoThe etiology of steroid cataractrdquo Journal of OcularPharmacology and Therapeutics vol 23 no 5 pp 403ndash4202007

[34] P Greaves Histopathological of Preclinical Toxicity StudiesElsevier New York NY USA 3rd edition 2007

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Behavioural Neurology

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Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Oxidative Medicine and Cellular Longevity 5

(1A) (2A) (3A) (4A) (5A)

(1B) (2B) (3B) (4B) (5B)

(1a) (2a) (3a) (4a) (5a)

(1b) (2b) (3b) (4b) (5b)

Figure 2 Representative photographs showing the effects of 10-week treatment of PWCG in prevention of cataract A photographs via slitlamp B photographs via camera 1 control rat 2 diabetic rat which received vehicle (DM + vehicle) 3 diabetic rat which received PWCGat dose of 50mgsdotkgminus1 BW (DM + PWCG 50mgsdotkgminus1 BW) 4 diabetic rat which received PWCG at dose of 100mgsdotkgminus1 BW (DM + PWCG100mgsdotkgminus1 BW) 5 diabetic rat which received PWCG at dose of 200mgsdotkgminus1 BW (DM + PWCG 200mgsdotkgminus1 BW)

33 The Effect of PWCG on Retinopathy in Diabetic Rats Theeffect of PWCG on histopathological change of retina wasshown in Figure 5 At 10-week study period the total retinalthickness (TRT) the thickness of the retinal outer nuclearlayer (ROT) and the number of cells in the ganglion celllayer (NG) of diabetic ratswhich received vehicle significantlydecreasedwhen compared to controlHowever the decreasedTRT ROT and NG induced by diabetic condition wereattenuated in diabetic rats which received all doses of PWCGFigures 6 7 and 8 showed that diabetic rats which receivedPWCG at doses of 50 100 and 200mgsdotkgminus1 BW significantlyincreased the TRT (119875 value lt 0001 all compared withdiabetic rats which received vehicle) ROT (119875 value lt 0001all compared with diabetic rats which received vehicle) andNG of retina (119875 value lt 0001 005 and 0001 resp comparedwith diabetic rats which received vehicle)

34 Effect of PWCG on Oxidative Stress Markers Based onthe crucial role of oxidative stress on cataract and retinopathythe effects of PWCG on oxidative stress markers includinglevel of MDA and the activities of SOD CAT and GPx inlens were carried out The results were shown in Figures 9

10 11 and 12 It was found that diabetic rats which receivedvehicle significantly increased MDA level (119875 value lt 0001compared with control) but decreased SOD CAT and GPxactivities (119875 value lt 0001 all compared with control) in lensPWCG at doses of 50 100 and 200mgsdotkgminus1 BW significantlymitigated the elevation of MDA level induced by diabetesmellitus (119875 value lt 0001 all compared with diabetic ratswhich received vehicle) Low dose of PWCG also mitigatedthe reduction of GPx activity in lens of diabetic rats (119875value lt 005 compared with diabetic rats which receivedvehicle) whereas high dose of PWCGmitigated the reductionof both CAT and GPx activities of lens of diabetic rats (119875value lt 001 and 0001 resp compared with diabetic ratswhich received vehicle) However no significant changes ofscavenger enzymes activities were observed in the diabeticrats which received medium dose of PWCG

35 Effect of PWCG on Aldose Reductase Activity In additionto the oxidative stress the polyol pathway also plays thecrucial role on the cataractogenesis Therefore the effectof PWCG on aldose reductase enzyme activity in lenswas also investigated and data were shown in Figure 13 It

6 Oxidative Medicine and Cellular Longevity

50 120583m

(a)

50 120583m

(b)

50 120583m

(c)

50 120583m

(d)

50 120583m

(e)

Figure 3 Photographs of transverse sections of eye balls (5 120583m thickness) stained with HampE which determined the cataract severity at theend of experiment using light microscope (week 10) (a) Control (b) DM + Vehicle (c) DM + PWCG (50mgkgsdotBW) (d) DM + PWCG(100mgkgsdotBW) and (e) DM + PWCG (200mgkgsdotBW)

000

050

100

150

200

250

300

350

400

Gra

de o

f cat

arac

t sev

erity

lowastlowastlowast

lowastlowastlowast

lowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 4 The cataract severity of control and diabetic rats (DM)which received either vehicle or PWCG at doses of 50 100 and200mgsdotkgminus1 BW for 10 weeks

119875 value lt 0001 compared withcontrol and lowastlowastlowast119875 value lt 0001 compared with vehicle

was demonstrated that aldose reductase activity in lens ofdiabetic rats was increased (119875 value lt 005 compared withcontrol) PWCG at dose of 200mgsdotkgminus1 BW could signifi-cantly decrease the elevation of aldose reductase induced bydiabetes mellitus in rat lens No other significant changeswere observed

4 Discussion

Diabetic cataract and retinopathy are the important causes ofblindness so the prevention of these conditions is very muchimportant Our results clearly demonstrated that PWCG sig-nificantly mitigated the reduction of ganglia in outer nuclearlayer (ONL) OUL thickness and total retina thickness Inaddition PWCG also showed anticataract effect Togetherwith the changes mentioned earlier the decreased oxidativestress and aldose reductase activity in rat lens were alsoobserved

It has been reported that the neurodegeneration of gan-glion cells plays the crucial role on the reduction of OULand total retina thickness [21] The degeneration of retinalganglion cells is under the influence of many factors suchas inflammation oxidative stress or exposure to advancedglycation end products [22] and polyol pathway [23] Ourdata also demonstrates the increased oxidative stress markerssuch asMDA level and the enhanced aldose reductase activityin lens of diabetic rats These changes are in agreement withthe previous study [22 23] Therefore we suggested thatthe effect of PWCG to attenuate retinopathy induced bydiabetes mellitus might be partly related to the decreasedoxidative stress and aldose reductase activity in rat lens Itwas found that the enhanced of GPx activity in rat lens mightbe responsible in part for the reduction of oxidative stress inrat lens of diabetic rats especially at low and high doses ofPWCG In addition to the increased GPx activity the highdose of PWCG also enhanced CAT in rat lens Therefore

Oxidative Medicine and Cellular Longevity 7

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(a)

50 120583m

RODS and CONES

ONL

OPLINL

IPLGCL

(b)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(c)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(d)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(e)

Figure 5 Light microscope of transverse section of eye ball (5 120583m thickness) stained with HampE which determined the pathological changeof retina at the end of experiment (week 10) (a) Control rat (b) diabetic rat which received vehicle (DM + vehicle) (c) diabetic rat whichreceived PWCG at dose of 50mgsdotkgminus1 BW (DM + PWCG 50mgsdotkgminus1 BW) (d) diabetic rat which received PWCG at dose of 100mgsdotkgminus1 BW(DM + PWCG 100mgsdotkgminus1 BW) (e) diabetic rat which received PWCG at dose of 200mgsdotkgminus1 BW (DM + PWCG 200mgsdotkgminus1 BW)

the elevation of both CAT and GPx might be responsible forthe decreased oxidative stress which in turn enhanced thesurvival of ganglion cells in OUL leading to the increasedtotal retina thickness However the decreased oxidative stressin rat lens induced by medium dose of PWCG might beassociated with other mechanism in addition to the elevationof the scavenger enzymes activities Since the excess oxidativestress occurs as the result of imbalance between oxidativestress formation and oxidative stress buffering capacities wesuggested that the decreased oxidative stress formationmightplay a role on the reduction of oxidative stress induced by thePWCG especially at the medium concentration

Oxidative stress and aldose reductase also play the crucialrole on the cataractogenesis in diabetic rats [24 25] In addi-tion it has been reported that antioxidant and aldose reduc-tase inhibitory effects can improve cataract [26] Thus theanticataractogenis of PWCG might also be associated withboth effects mentioned earlier Aldose reductase also playsa role on the pathophysiology of diabetic retinopathy [27]

Based on the basis of this information we do suggest thatthe effect of PWCG to improve retinopathy especially at highdose should also relate with the aldose reductase inhibitoryeffect

Previous study has demonstrated that flavonoids canprotect against oxidative stress and increase the survivalof ganglion cells [28] and prevent cataract [29] in diabeticrats Recent study has demonstrated that anthocyanins andanthocyanins-rich substance have the potential to protectagainst retinopathy [30] and diabetic cataract [26] In addi-tion ginger and its constituents also show the potential toprotect against diabetic retinopathy [31] Therefore the anti-cataractogenesis and antiretinopathy of PWCGmay be due tothe flavonoids such as anthocyanins in PWCG Other typesof flavonoidmay also play a role However further researchesare required to confirm the possible active ingredient

In this study the slit lamp examination clearly providedthe changes of lens characteristic However no pupil wasdilated during the grading of severity as that was performed

8 Oxidative Medicine and Cellular Longevity

0002000400060008000

100001200014000160001800020000

lowastlowastlowastlowastlowastlowast

lowastlowastlowast

Tota

l ret

inal

thic

knes

s (120583

m)

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 6 The total retinal thickness of retina (TRT) of control anddiabetic rats (DM) which received either vehicle or the combinationextract of purple waxy corn and ginger (PWCG) at doses of 50 100and 200mgsdotkgminus1 BW for 10weeks 119875 valuelt 0001 comparedwithcontrol and lowastlowastlowast119875 value lt 0001 compared with vehicle

000500

100015002000250030003500400045005000

The r

etin

al n

ucle

ar la

yer t

hick

ness

(120583m

)

lowastlowastlowast lowastlowastlowastlowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 7 The thickness of the retinal outer nuclear layer (ROT)of control and diabetic rats (DM) which received either vehicle orthe combination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks

119875 valuelt 0001 compared with control and lowastlowastlowast119875 value lt 0001 comparedwith vehicle

during the slit lamp examination in humanThismay possiblyinfluence the precision of grading scores which reflect thereal severity of cataract To obtain the precise severity ofcataract the slit lamp examination should be performedwhile pupil was dilated However all rats were assessed underthe condition without the pupil dilator so no confoundingerror about the beneficial effect of PWCG on cataract wasachieved

The lenticular opacification in control rats was alsoobserved during the 6ndash10 weeks of study period Based onthe previous study that stress hormone or glucocorticoids

000500

100015002000250030003500400045005000

The n

umbe

r of n

euro

nin

gan

glio

n ce

ll lay

er in

retin

a

lowastlowastlowastlowastlowastlowast

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 8 The number of cells in the ganglion cell layer (NG)in retina of control and diabetic rats (DM) which received eithervehicle or the combination extract of purple waxy corn and ginger(PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks 119875value lt 0001 compared with control and lowast lowastlowastlowast119875 value lt 005 and0001 compared with vehicle

MD

A (U

mg

prot

ein)

000

100

200

300

400

500

600

lowastlowastlowast lowastlowastlowastlowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 9 The level of malondialdehyde (MDA) (Umg protein) inlens of control and diabetic rats (DM) which received either vehicleor the combination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 = 8group)119875 value lt 0001 compared with control and lowastlowastlowast119875 value lt 0001

compared with diabetic rats which received vehicle (DM + vehicle)

can also induce cataract [32 33] we do suggest that thedevelopment of cataract in control group may possibly occuras the exposure to stress condition leading to the elevationof glucocorticoids hormone resulting in the cataractogenesisUnfortunately the level of corticosterone has not been deter-mined in this study Therefore the precise understandingmechanism of this change requires further investigation Inaddition spontaneous age-related cataract can also occurin laboratory rodents The prevalence is varied dependingon species Even young adolescent rodents can be found tohave cataract approximately 20 [34]Therefore the cataract

Oxidative Medicine and Cellular Longevity 9SO

D (U

mg

prot

ein)

000050100150200250300350400450

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 10 The activity of superoxide dismutase (SOD) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BWfor 10weeks(119873 = 8group) 119875 value lt 0001 compared with control

CAT

(Um

g pr

otei

n)

000

1000

2000

3000

4000

5000

6000

lowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 11 The activity of catalase (CAT) (Umg protein) in lensof control and diabetic rats (DM) which received either vehicle orthe combination extract of purple waxy corn and ginger (PWCG) atdoses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 = 8group)119875 value lt 0001 compared with control and lowastlowast119875 value lt 001

compared with diabetic rats which received vehicle (DM + vehicle)

observed in normal rats might occur either via stress orvia spontaneous cataractogenesis of the laboratory rodentsmentioned earlier

5 Conclusion

The combination extract of purple waxy corn seeds andginger rhizome (PWCG) can protect against diabetic cataractand diabetic retinopathyThepossible underlyingmechanismmay occur partly via the decreased oxidative stress and thesuppression of aldose reductase activity in lens The possibleactive ingredient may be associated with flavonoids such as

000020040060080100120140160180200

GPx

(Um

g pr

otei

n)

lowastlowastlowast

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 12 The activity of glutathione peroxidase (GPx) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10weeks (119873 = 8group) 119875 value lt 0001 compared with controland lowast lowastlowastlowast119875 value lt 005 0001 respectively compared with diabeticrats which received vehicle (DM + vehicle)

AR

activ

ity (n

mol

min

mg)

00000

00010

00020

00030

00040

00050

00060

00070

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 13 The activity of aldose reductase (AR) (nmolminmg)in lens of control and diabetic rats (DM) which received eithervehicle or the combination extract of purple waxy corn and ginger(PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 =8group) 119875 value lt 005 compared with control and lowast119875 value lt005 compared with diabetic rats which received vehicle (DM +vehicle)

anthocyanin and other flavonoids Further researches are stillessential to understand the precise underlying mechanismand possible active ingredient

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

10 Oxidative Medicine and Cellular Longevity

Acknowledgments

This study was supported by the Higher Education ResearchPromotion andNational Research University Project ofThai-land Office of the Higher Education Commission throughthe Food and Functional Food Research Cluster of KhonKaen University and the Integrative Complementary Alter-native Medicine Research and Development Center KhonKaen University Thailand Great thanks were also extendedto Research and Development Center of Herbal HealthProduct for providing the opportunity to collaborate in thesubcluster

References

[1] R L Engerman and T S Kern ldquoHyperglycemia as a cause ofdiabetic retinopathyrdquo Metabolism vol 35 supplement 1 no 4pp 20ndash23 1986

[2] A Pollreisz and U Schmidt-Erfurth ldquoDiabetic cataractmdashpathogenesis epidemiology and treatmentrdquo Journal of Ophthal-mology vol 2010 Article ID 608751 8 pages 2010

[3] I G Obrosova S S M Chung and P F Kador ldquoDiabeticcataracts mechanisms and managementrdquo DiabetesMetabolismResearch and Reviews vol 26 no 3 pp 172ndash180 2010

[4] J Ding and T Y Wong ldquoCurrent epidemiology of diabeticretinopathy and diabetic macular edemardquo Current DiabetesReports vol 12 no 4 pp 346ndash354 2012

[5] A Y W Lee and S S M Chung ldquoContributions of polyolpathway to oxidative stress in diabetic cataractrdquo The FASEBJournal vol 13 no 1 pp 23ndash30 1999

[6] S A Madsen-Bouterse and R A Kowluru ldquoOxidative stressand diabetic retinopathy pathophysiological mechanisms andtreatment perspectivesrdquo Reviews in Endocrine and MetabolicDisorders vol 9 no 4 pp 315ndash327 2008

[7] I G Obrosova and P F Kador ldquoAldose reductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 1389ndash2010 2011

[8] V R Drel P Pacher T K Ali et al ldquoAldose reductase inhibitorfidarestat counteracts diabetes-associated cataract formationretinal oxidative-nitrosative stress glial activation and apopto-sisrdquo International Journal of Molecular Medicine vol 21 no 6pp 667ndash676 2008

[9] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (Purple Waxy Corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[10] Y Morimitsu K Kubota T Tashiro E Hashizume T Kamiyaand T Osawa ldquoInhibitory effect of anthocyanins and coloredrice ondiabetic cataract formation in the rat lensesrdquo Interna-tional Congress Series vol 1245 pp 503ndash508 2002

[11] M Saraswat P Suryanarayana P Y Reddy M A Patil NBalakrishna and G B Reddy ldquoAntiglycating potential of Zin-giber officinalis and delay of diabetic cataract in ratsrdquoMolecularVision vol 16 pp 1525ndash1537 2010

[12] Y Li V H Tran C C Duke and B D Roufogalis ldquoPreventiveand protective properties of Zingiber officinale (Ginger) indiabetes mellitus diabetic complications and associated lipidand other metabolic disorders a brief reviewrdquo Evidence-basedComplementary and Alternative Medicine vol 2012 Article ID516870 10 pages 2012

[13] P Suryanarayana M Saraswat T Mrudula T P Krishna KKrishnaswamy andG B Reddy ldquoCurcumin and turmeric delaystreptozotocin-induced diabetic cataract in ratsrdquo InvestigativeOphthalmology amp Visual Science vol 46 no 6 pp 2092ndash20992005

[14] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[15] Y Sun L W Oberley and Y Li ldquoA simple method for clinicalassay of superoxide dismutaserdquo Clinical Chemistry vol 34 no3 pp 497ndash500 1988

[16] L Goth ldquoA simple method for determination of serum catalaseactivity and revision of reference rangerdquo Clinica Chimica Actavol 196 no 2-3 pp 143ndash151 1991

[17] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973

[18] M B Patel and S M Mishra ldquoAldose reductase inhibitoryactivity and anti catraract potential of some traditionallyacclaimed antidiabetic medicinal plantsrdquo Oriental Pharmacyand Experimental Medicine vol 9 no 3 pp 245ndash251 2009

[19] R Agarwal I Iezhitsa N A Awaludin et al ldquoEffects ofmagnesium taurate on the onset and progression of galactose-induced experimental cataract in vivo and in vitro evaluationrdquoExperimental Eye Research vol 110 pp 35ndash43 2013

[20] P M Martin P Roon T K van Ells V Ganapathy and SB Smith ldquoDeath of retinal neurons in streptozotocin-induceddiabetic micerdquo Investigative Ophthalmology and Visual Sciencevol 45 no 9 pp 3330ndash3336 2004

[21] H W van Dijk F D Verbraak P H B Kok et al ldquoDecreasedretinal ganglion cell layer thickness in patients with type 1diabetesrdquo Investigative Ophthalmology and Visual Science vol51 no 7 pp 3660ndash3665 2010

[22] T S Kern and A J Barber ldquoRetinal ganglion cells in diabetesrdquoJournal of Physiology vol 586 no 18 pp 4401ndash4408 2008

[23] G A Ramadan ldquoSorbitol-induced diabetic-like retinal lesionsin rats microscopic studyrdquo American Journal of Pharmacologyand Toxicology vol 2 no 2 pp 89ndash97 2007

[24] B Ozmen D Ozmen E Erkin I Guner S Habif and OBaymdir ldquoLens superoxide dismutase and catalase activities indiabetic cataractrdquoClinical Biochemistry vol 35 no 1 pp 69ndash722002

[25] A B M Reddy R Tammali R Mishra S Srivastava S KSrivastava and K V Ramana ldquoAldose reductase deficiencyprotects sugar-induced lens opacification in ratsrdquo Chemico-Biological Interactions vol 191 no 1ndash3 pp 346ndash350 2011

[26] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (purple waxy corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[27] I G Obrosova and P F Kador ldquoAldosereductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 373ndash385 2011

[28] PMaher andAHanneken ldquoFlavonoids protect retinal ganglioncells from oxidative stress-induced deathrdquo Investigative Oph-thalmology and Visual Science vol 46 no 12 pp 4796ndash48032005

[29] M Stefek ldquoNatural flavonoids as potential multifunctionalagents in prevention of diabetic cataractrdquo InterdisciplinaryToxicology vol 4 no 2 pp 69ndash77 2011

Oxidative Medicine and Cellular Longevity 11

[30] S F Nabavi S Habtemariam M Daglia N Shafighi A JBarber and S M Nabavi ldquoAnthocyanins as a potential therapyfor diabetic retinopathyrdquo Current Medicinal Chemistry vol 22no 1 pp 51ndash58 2014

[31] A H Rahmani F M Al Shabrmi and S M Aly ldquoActiveingredients of ginger as potential candidates in the preventionand treatment of diseases via modulation of biological activ-itiesrdquo International Journal of Physiology Pathophysiology andPharmacology vol 6 no 2 pp 125ndash136 2014

[32] H Watanabe H Kosano and H Nishigori ldquoSteroid-inducedshort term diabetes in chick embryo reversible effects of insulinon metabolic changes and cataract formationrdquo InvestigativeOphthalmology amp Visual Science vol 41 no 7 pp 1846ndash18522000

[33] E R James ldquoThe etiology of steroid cataractrdquo Journal of OcularPharmacology and Therapeutics vol 23 no 5 pp 403ndash4202007

[34] P Greaves Histopathological of Preclinical Toxicity StudiesElsevier New York NY USA 3rd edition 2007

Submit your manuscripts athttpwwwhindawicom

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

6 Oxidative Medicine and Cellular Longevity

50 120583m

(a)

50 120583m

(b)

50 120583m

(c)

50 120583m

(d)

50 120583m

(e)

Figure 3 Photographs of transverse sections of eye balls (5 120583m thickness) stained with HampE which determined the cataract severity at theend of experiment using light microscope (week 10) (a) Control (b) DM + Vehicle (c) DM + PWCG (50mgkgsdotBW) (d) DM + PWCG(100mgkgsdotBW) and (e) DM + PWCG (200mgkgsdotBW)

000

050

100

150

200

250

300

350

400

Gra

de o

f cat

arac

t sev

erity

lowastlowastlowast

lowastlowastlowast

lowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 4 The cataract severity of control and diabetic rats (DM)which received either vehicle or PWCG at doses of 50 100 and200mgsdotkgminus1 BW for 10 weeks

119875 value lt 0001 compared withcontrol and lowastlowastlowast119875 value lt 0001 compared with vehicle

was demonstrated that aldose reductase activity in lens ofdiabetic rats was increased (119875 value lt 005 compared withcontrol) PWCG at dose of 200mgsdotkgminus1 BW could signifi-cantly decrease the elevation of aldose reductase induced bydiabetes mellitus in rat lens No other significant changeswere observed

4 Discussion

Diabetic cataract and retinopathy are the important causes ofblindness so the prevention of these conditions is very muchimportant Our results clearly demonstrated that PWCG sig-nificantly mitigated the reduction of ganglia in outer nuclearlayer (ONL) OUL thickness and total retina thickness Inaddition PWCG also showed anticataract effect Togetherwith the changes mentioned earlier the decreased oxidativestress and aldose reductase activity in rat lens were alsoobserved

It has been reported that the neurodegeneration of gan-glion cells plays the crucial role on the reduction of OULand total retina thickness [21] The degeneration of retinalganglion cells is under the influence of many factors suchas inflammation oxidative stress or exposure to advancedglycation end products [22] and polyol pathway [23] Ourdata also demonstrates the increased oxidative stress markerssuch asMDA level and the enhanced aldose reductase activityin lens of diabetic rats These changes are in agreement withthe previous study [22 23] Therefore we suggested thatthe effect of PWCG to attenuate retinopathy induced bydiabetes mellitus might be partly related to the decreasedoxidative stress and aldose reductase activity in rat lens Itwas found that the enhanced of GPx activity in rat lens mightbe responsible in part for the reduction of oxidative stress inrat lens of diabetic rats especially at low and high doses ofPWCG In addition to the increased GPx activity the highdose of PWCG also enhanced CAT in rat lens Therefore

Oxidative Medicine and Cellular Longevity 7

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(a)

50 120583m

RODS and CONES

ONL

OPLINL

IPLGCL

(b)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(c)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(d)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(e)

Figure 5 Light microscope of transverse section of eye ball (5 120583m thickness) stained with HampE which determined the pathological changeof retina at the end of experiment (week 10) (a) Control rat (b) diabetic rat which received vehicle (DM + vehicle) (c) diabetic rat whichreceived PWCG at dose of 50mgsdotkgminus1 BW (DM + PWCG 50mgsdotkgminus1 BW) (d) diabetic rat which received PWCG at dose of 100mgsdotkgminus1 BW(DM + PWCG 100mgsdotkgminus1 BW) (e) diabetic rat which received PWCG at dose of 200mgsdotkgminus1 BW (DM + PWCG 200mgsdotkgminus1 BW)

the elevation of both CAT and GPx might be responsible forthe decreased oxidative stress which in turn enhanced thesurvival of ganglion cells in OUL leading to the increasedtotal retina thickness However the decreased oxidative stressin rat lens induced by medium dose of PWCG might beassociated with other mechanism in addition to the elevationof the scavenger enzymes activities Since the excess oxidativestress occurs as the result of imbalance between oxidativestress formation and oxidative stress buffering capacities wesuggested that the decreased oxidative stress formationmightplay a role on the reduction of oxidative stress induced by thePWCG especially at the medium concentration

Oxidative stress and aldose reductase also play the crucialrole on the cataractogenesis in diabetic rats [24 25] In addi-tion it has been reported that antioxidant and aldose reduc-tase inhibitory effects can improve cataract [26] Thus theanticataractogenis of PWCG might also be associated withboth effects mentioned earlier Aldose reductase also playsa role on the pathophysiology of diabetic retinopathy [27]

Based on the basis of this information we do suggest thatthe effect of PWCG to improve retinopathy especially at highdose should also relate with the aldose reductase inhibitoryeffect

Previous study has demonstrated that flavonoids canprotect against oxidative stress and increase the survivalof ganglion cells [28] and prevent cataract [29] in diabeticrats Recent study has demonstrated that anthocyanins andanthocyanins-rich substance have the potential to protectagainst retinopathy [30] and diabetic cataract [26] In addi-tion ginger and its constituents also show the potential toprotect against diabetic retinopathy [31] Therefore the anti-cataractogenesis and antiretinopathy of PWCGmay be due tothe flavonoids such as anthocyanins in PWCG Other typesof flavonoidmay also play a role However further researchesare required to confirm the possible active ingredient

In this study the slit lamp examination clearly providedthe changes of lens characteristic However no pupil wasdilated during the grading of severity as that was performed

8 Oxidative Medicine and Cellular Longevity

0002000400060008000

100001200014000160001800020000

lowastlowastlowastlowastlowastlowast

lowastlowastlowast

Tota

l ret

inal

thic

knes

s (120583

m)

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 6 The total retinal thickness of retina (TRT) of control anddiabetic rats (DM) which received either vehicle or the combinationextract of purple waxy corn and ginger (PWCG) at doses of 50 100and 200mgsdotkgminus1 BW for 10weeks 119875 valuelt 0001 comparedwithcontrol and lowastlowastlowast119875 value lt 0001 compared with vehicle

000500

100015002000250030003500400045005000

The r

etin

al n

ucle

ar la

yer t

hick

ness

(120583m

)

lowastlowastlowast lowastlowastlowastlowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 7 The thickness of the retinal outer nuclear layer (ROT)of control and diabetic rats (DM) which received either vehicle orthe combination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks

119875 valuelt 0001 compared with control and lowastlowastlowast119875 value lt 0001 comparedwith vehicle

during the slit lamp examination in humanThismay possiblyinfluence the precision of grading scores which reflect thereal severity of cataract To obtain the precise severity ofcataract the slit lamp examination should be performedwhile pupil was dilated However all rats were assessed underthe condition without the pupil dilator so no confoundingerror about the beneficial effect of PWCG on cataract wasachieved

The lenticular opacification in control rats was alsoobserved during the 6ndash10 weeks of study period Based onthe previous study that stress hormone or glucocorticoids

000500

100015002000250030003500400045005000

The n

umbe

r of n

euro

nin

gan

glio

n ce

ll lay

er in

retin

a

lowastlowastlowastlowastlowastlowast

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 8 The number of cells in the ganglion cell layer (NG)in retina of control and diabetic rats (DM) which received eithervehicle or the combination extract of purple waxy corn and ginger(PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks 119875value lt 0001 compared with control and lowast lowastlowastlowast119875 value lt 005 and0001 compared with vehicle

MD

A (U

mg

prot

ein)

000

100

200

300

400

500

600

lowastlowastlowast lowastlowastlowastlowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 9 The level of malondialdehyde (MDA) (Umg protein) inlens of control and diabetic rats (DM) which received either vehicleor the combination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 = 8group)119875 value lt 0001 compared with control and lowastlowastlowast119875 value lt 0001

compared with diabetic rats which received vehicle (DM + vehicle)

can also induce cataract [32 33] we do suggest that thedevelopment of cataract in control group may possibly occuras the exposure to stress condition leading to the elevationof glucocorticoids hormone resulting in the cataractogenesisUnfortunately the level of corticosterone has not been deter-mined in this study Therefore the precise understandingmechanism of this change requires further investigation Inaddition spontaneous age-related cataract can also occurin laboratory rodents The prevalence is varied dependingon species Even young adolescent rodents can be found tohave cataract approximately 20 [34]Therefore the cataract

Oxidative Medicine and Cellular Longevity 9SO

D (U

mg

prot

ein)

000050100150200250300350400450

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 10 The activity of superoxide dismutase (SOD) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BWfor 10weeks(119873 = 8group) 119875 value lt 0001 compared with control

CAT

(Um

g pr

otei

n)

000

1000

2000

3000

4000

5000

6000

lowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 11 The activity of catalase (CAT) (Umg protein) in lensof control and diabetic rats (DM) which received either vehicle orthe combination extract of purple waxy corn and ginger (PWCG) atdoses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 = 8group)119875 value lt 0001 compared with control and lowastlowast119875 value lt 001

compared with diabetic rats which received vehicle (DM + vehicle)

observed in normal rats might occur either via stress orvia spontaneous cataractogenesis of the laboratory rodentsmentioned earlier

5 Conclusion

The combination extract of purple waxy corn seeds andginger rhizome (PWCG) can protect against diabetic cataractand diabetic retinopathyThepossible underlyingmechanismmay occur partly via the decreased oxidative stress and thesuppression of aldose reductase activity in lens The possibleactive ingredient may be associated with flavonoids such as

000020040060080100120140160180200

GPx

(Um

g pr

otei

n)

lowastlowastlowast

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 12 The activity of glutathione peroxidase (GPx) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10weeks (119873 = 8group) 119875 value lt 0001 compared with controland lowast lowastlowastlowast119875 value lt 005 0001 respectively compared with diabeticrats which received vehicle (DM + vehicle)

AR

activ

ity (n

mol

min

mg)

00000

00010

00020

00030

00040

00050

00060

00070

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 13 The activity of aldose reductase (AR) (nmolminmg)in lens of control and diabetic rats (DM) which received eithervehicle or the combination extract of purple waxy corn and ginger(PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 =8group) 119875 value lt 005 compared with control and lowast119875 value lt005 compared with diabetic rats which received vehicle (DM +vehicle)

anthocyanin and other flavonoids Further researches are stillessential to understand the precise underlying mechanismand possible active ingredient

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

10 Oxidative Medicine and Cellular Longevity

Acknowledgments

This study was supported by the Higher Education ResearchPromotion andNational Research University Project ofThai-land Office of the Higher Education Commission throughthe Food and Functional Food Research Cluster of KhonKaen University and the Integrative Complementary Alter-native Medicine Research and Development Center KhonKaen University Thailand Great thanks were also extendedto Research and Development Center of Herbal HealthProduct for providing the opportunity to collaborate in thesubcluster

References

[1] R L Engerman and T S Kern ldquoHyperglycemia as a cause ofdiabetic retinopathyrdquo Metabolism vol 35 supplement 1 no 4pp 20ndash23 1986

[2] A Pollreisz and U Schmidt-Erfurth ldquoDiabetic cataractmdashpathogenesis epidemiology and treatmentrdquo Journal of Ophthal-mology vol 2010 Article ID 608751 8 pages 2010

[3] I G Obrosova S S M Chung and P F Kador ldquoDiabeticcataracts mechanisms and managementrdquo DiabetesMetabolismResearch and Reviews vol 26 no 3 pp 172ndash180 2010

[4] J Ding and T Y Wong ldquoCurrent epidemiology of diabeticretinopathy and diabetic macular edemardquo Current DiabetesReports vol 12 no 4 pp 346ndash354 2012

[5] A Y W Lee and S S M Chung ldquoContributions of polyolpathway to oxidative stress in diabetic cataractrdquo The FASEBJournal vol 13 no 1 pp 23ndash30 1999

[6] S A Madsen-Bouterse and R A Kowluru ldquoOxidative stressand diabetic retinopathy pathophysiological mechanisms andtreatment perspectivesrdquo Reviews in Endocrine and MetabolicDisorders vol 9 no 4 pp 315ndash327 2008

[7] I G Obrosova and P F Kador ldquoAldose reductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 1389ndash2010 2011

[8] V R Drel P Pacher T K Ali et al ldquoAldose reductase inhibitorfidarestat counteracts diabetes-associated cataract formationretinal oxidative-nitrosative stress glial activation and apopto-sisrdquo International Journal of Molecular Medicine vol 21 no 6pp 667ndash676 2008

[9] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (Purple Waxy Corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[10] Y Morimitsu K Kubota T Tashiro E Hashizume T Kamiyaand T Osawa ldquoInhibitory effect of anthocyanins and coloredrice ondiabetic cataract formation in the rat lensesrdquo Interna-tional Congress Series vol 1245 pp 503ndash508 2002

[11] M Saraswat P Suryanarayana P Y Reddy M A Patil NBalakrishna and G B Reddy ldquoAntiglycating potential of Zin-giber officinalis and delay of diabetic cataract in ratsrdquoMolecularVision vol 16 pp 1525ndash1537 2010

[12] Y Li V H Tran C C Duke and B D Roufogalis ldquoPreventiveand protective properties of Zingiber officinale (Ginger) indiabetes mellitus diabetic complications and associated lipidand other metabolic disorders a brief reviewrdquo Evidence-basedComplementary and Alternative Medicine vol 2012 Article ID516870 10 pages 2012

[13] P Suryanarayana M Saraswat T Mrudula T P Krishna KKrishnaswamy andG B Reddy ldquoCurcumin and turmeric delaystreptozotocin-induced diabetic cataract in ratsrdquo InvestigativeOphthalmology amp Visual Science vol 46 no 6 pp 2092ndash20992005

[14] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[15] Y Sun L W Oberley and Y Li ldquoA simple method for clinicalassay of superoxide dismutaserdquo Clinical Chemistry vol 34 no3 pp 497ndash500 1988

[16] L Goth ldquoA simple method for determination of serum catalaseactivity and revision of reference rangerdquo Clinica Chimica Actavol 196 no 2-3 pp 143ndash151 1991

[17] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973

[18] M B Patel and S M Mishra ldquoAldose reductase inhibitoryactivity and anti catraract potential of some traditionallyacclaimed antidiabetic medicinal plantsrdquo Oriental Pharmacyand Experimental Medicine vol 9 no 3 pp 245ndash251 2009

[19] R Agarwal I Iezhitsa N A Awaludin et al ldquoEffects ofmagnesium taurate on the onset and progression of galactose-induced experimental cataract in vivo and in vitro evaluationrdquoExperimental Eye Research vol 110 pp 35ndash43 2013

[20] P M Martin P Roon T K van Ells V Ganapathy and SB Smith ldquoDeath of retinal neurons in streptozotocin-induceddiabetic micerdquo Investigative Ophthalmology and Visual Sciencevol 45 no 9 pp 3330ndash3336 2004

[21] H W van Dijk F D Verbraak P H B Kok et al ldquoDecreasedretinal ganglion cell layer thickness in patients with type 1diabetesrdquo Investigative Ophthalmology and Visual Science vol51 no 7 pp 3660ndash3665 2010

[22] T S Kern and A J Barber ldquoRetinal ganglion cells in diabetesrdquoJournal of Physiology vol 586 no 18 pp 4401ndash4408 2008

[23] G A Ramadan ldquoSorbitol-induced diabetic-like retinal lesionsin rats microscopic studyrdquo American Journal of Pharmacologyand Toxicology vol 2 no 2 pp 89ndash97 2007

[24] B Ozmen D Ozmen E Erkin I Guner S Habif and OBaymdir ldquoLens superoxide dismutase and catalase activities indiabetic cataractrdquoClinical Biochemistry vol 35 no 1 pp 69ndash722002

[25] A B M Reddy R Tammali R Mishra S Srivastava S KSrivastava and K V Ramana ldquoAldose reductase deficiencyprotects sugar-induced lens opacification in ratsrdquo Chemico-Biological Interactions vol 191 no 1ndash3 pp 346ndash350 2011

[26] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (purple waxy corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[27] I G Obrosova and P F Kador ldquoAldosereductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 373ndash385 2011

[28] PMaher andAHanneken ldquoFlavonoids protect retinal ganglioncells from oxidative stress-induced deathrdquo Investigative Oph-thalmology and Visual Science vol 46 no 12 pp 4796ndash48032005

[29] M Stefek ldquoNatural flavonoids as potential multifunctionalagents in prevention of diabetic cataractrdquo InterdisciplinaryToxicology vol 4 no 2 pp 69ndash77 2011

Oxidative Medicine and Cellular Longevity 11

[30] S F Nabavi S Habtemariam M Daglia N Shafighi A JBarber and S M Nabavi ldquoAnthocyanins as a potential therapyfor diabetic retinopathyrdquo Current Medicinal Chemistry vol 22no 1 pp 51ndash58 2014

[31] A H Rahmani F M Al Shabrmi and S M Aly ldquoActiveingredients of ginger as potential candidates in the preventionand treatment of diseases via modulation of biological activ-itiesrdquo International Journal of Physiology Pathophysiology andPharmacology vol 6 no 2 pp 125ndash136 2014

[32] H Watanabe H Kosano and H Nishigori ldquoSteroid-inducedshort term diabetes in chick embryo reversible effects of insulinon metabolic changes and cataract formationrdquo InvestigativeOphthalmology amp Visual Science vol 41 no 7 pp 1846ndash18522000

[33] E R James ldquoThe etiology of steroid cataractrdquo Journal of OcularPharmacology and Therapeutics vol 23 no 5 pp 403ndash4202007

[34] P Greaves Histopathological of Preclinical Toxicity StudiesElsevier New York NY USA 3rd edition 2007

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Oxidative Medicine and Cellular Longevity 7

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(a)

50 120583m

RODS and CONES

ONL

OPLINL

IPLGCL

(b)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(c)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(d)

50 120583m

RODS and CONES

ONL

OPL

INL

IPL

GCL

(e)

Figure 5 Light microscope of transverse section of eye ball (5 120583m thickness) stained with HampE which determined the pathological changeof retina at the end of experiment (week 10) (a) Control rat (b) diabetic rat which received vehicle (DM + vehicle) (c) diabetic rat whichreceived PWCG at dose of 50mgsdotkgminus1 BW (DM + PWCG 50mgsdotkgminus1 BW) (d) diabetic rat which received PWCG at dose of 100mgsdotkgminus1 BW(DM + PWCG 100mgsdotkgminus1 BW) (e) diabetic rat which received PWCG at dose of 200mgsdotkgminus1 BW (DM + PWCG 200mgsdotkgminus1 BW)

the elevation of both CAT and GPx might be responsible forthe decreased oxidative stress which in turn enhanced thesurvival of ganglion cells in OUL leading to the increasedtotal retina thickness However the decreased oxidative stressin rat lens induced by medium dose of PWCG might beassociated with other mechanism in addition to the elevationof the scavenger enzymes activities Since the excess oxidativestress occurs as the result of imbalance between oxidativestress formation and oxidative stress buffering capacities wesuggested that the decreased oxidative stress formationmightplay a role on the reduction of oxidative stress induced by thePWCG especially at the medium concentration

Oxidative stress and aldose reductase also play the crucialrole on the cataractogenesis in diabetic rats [24 25] In addi-tion it has been reported that antioxidant and aldose reduc-tase inhibitory effects can improve cataract [26] Thus theanticataractogenis of PWCG might also be associated withboth effects mentioned earlier Aldose reductase also playsa role on the pathophysiology of diabetic retinopathy [27]

Based on the basis of this information we do suggest thatthe effect of PWCG to improve retinopathy especially at highdose should also relate with the aldose reductase inhibitoryeffect

Previous study has demonstrated that flavonoids canprotect against oxidative stress and increase the survivalof ganglion cells [28] and prevent cataract [29] in diabeticrats Recent study has demonstrated that anthocyanins andanthocyanins-rich substance have the potential to protectagainst retinopathy [30] and diabetic cataract [26] In addi-tion ginger and its constituents also show the potential toprotect against diabetic retinopathy [31] Therefore the anti-cataractogenesis and antiretinopathy of PWCGmay be due tothe flavonoids such as anthocyanins in PWCG Other typesof flavonoidmay also play a role However further researchesare required to confirm the possible active ingredient

In this study the slit lamp examination clearly providedthe changes of lens characteristic However no pupil wasdilated during the grading of severity as that was performed

8 Oxidative Medicine and Cellular Longevity

0002000400060008000

100001200014000160001800020000

lowastlowastlowastlowastlowastlowast

lowastlowastlowast

Tota

l ret

inal

thic

knes

s (120583

m)

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 6 The total retinal thickness of retina (TRT) of control anddiabetic rats (DM) which received either vehicle or the combinationextract of purple waxy corn and ginger (PWCG) at doses of 50 100and 200mgsdotkgminus1 BW for 10weeks 119875 valuelt 0001 comparedwithcontrol and lowastlowastlowast119875 value lt 0001 compared with vehicle

000500

100015002000250030003500400045005000

The r

etin

al n

ucle

ar la

yer t

hick

ness

(120583m

)

lowastlowastlowast lowastlowastlowastlowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 7 The thickness of the retinal outer nuclear layer (ROT)of control and diabetic rats (DM) which received either vehicle orthe combination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks

119875 valuelt 0001 compared with control and lowastlowastlowast119875 value lt 0001 comparedwith vehicle

during the slit lamp examination in humanThismay possiblyinfluence the precision of grading scores which reflect thereal severity of cataract To obtain the precise severity ofcataract the slit lamp examination should be performedwhile pupil was dilated However all rats were assessed underthe condition without the pupil dilator so no confoundingerror about the beneficial effect of PWCG on cataract wasachieved

The lenticular opacification in control rats was alsoobserved during the 6ndash10 weeks of study period Based onthe previous study that stress hormone or glucocorticoids

000500

100015002000250030003500400045005000

The n

umbe

r of n

euro

nin

gan

glio

n ce

ll lay

er in

retin

a

lowastlowastlowastlowastlowastlowast

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 8 The number of cells in the ganglion cell layer (NG)in retina of control and diabetic rats (DM) which received eithervehicle or the combination extract of purple waxy corn and ginger(PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks 119875value lt 0001 compared with control and lowast lowastlowastlowast119875 value lt 005 and0001 compared with vehicle

MD

A (U

mg

prot

ein)

000

100

200

300

400

500

600

lowastlowastlowast lowastlowastlowastlowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 9 The level of malondialdehyde (MDA) (Umg protein) inlens of control and diabetic rats (DM) which received either vehicleor the combination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 = 8group)119875 value lt 0001 compared with control and lowastlowastlowast119875 value lt 0001

compared with diabetic rats which received vehicle (DM + vehicle)

can also induce cataract [32 33] we do suggest that thedevelopment of cataract in control group may possibly occuras the exposure to stress condition leading to the elevationof glucocorticoids hormone resulting in the cataractogenesisUnfortunately the level of corticosterone has not been deter-mined in this study Therefore the precise understandingmechanism of this change requires further investigation Inaddition spontaneous age-related cataract can also occurin laboratory rodents The prevalence is varied dependingon species Even young adolescent rodents can be found tohave cataract approximately 20 [34]Therefore the cataract

Oxidative Medicine and Cellular Longevity 9SO

D (U

mg

prot

ein)

000050100150200250300350400450

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 10 The activity of superoxide dismutase (SOD) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BWfor 10weeks(119873 = 8group) 119875 value lt 0001 compared with control

CAT

(Um

g pr

otei

n)

000

1000

2000

3000

4000

5000

6000

lowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 11 The activity of catalase (CAT) (Umg protein) in lensof control and diabetic rats (DM) which received either vehicle orthe combination extract of purple waxy corn and ginger (PWCG) atdoses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 = 8group)119875 value lt 0001 compared with control and lowastlowast119875 value lt 001

compared with diabetic rats which received vehicle (DM + vehicle)

observed in normal rats might occur either via stress orvia spontaneous cataractogenesis of the laboratory rodentsmentioned earlier

5 Conclusion

The combination extract of purple waxy corn seeds andginger rhizome (PWCG) can protect against diabetic cataractand diabetic retinopathyThepossible underlyingmechanismmay occur partly via the decreased oxidative stress and thesuppression of aldose reductase activity in lens The possibleactive ingredient may be associated with flavonoids such as

000020040060080100120140160180200

GPx

(Um

g pr

otei

n)

lowastlowastlowast

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 12 The activity of glutathione peroxidase (GPx) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10weeks (119873 = 8group) 119875 value lt 0001 compared with controland lowast lowastlowastlowast119875 value lt 005 0001 respectively compared with diabeticrats which received vehicle (DM + vehicle)

AR

activ

ity (n

mol

min

mg)

00000

00010

00020

00030

00040

00050

00060

00070

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 13 The activity of aldose reductase (AR) (nmolminmg)in lens of control and diabetic rats (DM) which received eithervehicle or the combination extract of purple waxy corn and ginger(PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 =8group) 119875 value lt 005 compared with control and lowast119875 value lt005 compared with diabetic rats which received vehicle (DM +vehicle)

anthocyanin and other flavonoids Further researches are stillessential to understand the precise underlying mechanismand possible active ingredient

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

10 Oxidative Medicine and Cellular Longevity

Acknowledgments

This study was supported by the Higher Education ResearchPromotion andNational Research University Project ofThai-land Office of the Higher Education Commission throughthe Food and Functional Food Research Cluster of KhonKaen University and the Integrative Complementary Alter-native Medicine Research and Development Center KhonKaen University Thailand Great thanks were also extendedto Research and Development Center of Herbal HealthProduct for providing the opportunity to collaborate in thesubcluster

References

[1] R L Engerman and T S Kern ldquoHyperglycemia as a cause ofdiabetic retinopathyrdquo Metabolism vol 35 supplement 1 no 4pp 20ndash23 1986

[2] A Pollreisz and U Schmidt-Erfurth ldquoDiabetic cataractmdashpathogenesis epidemiology and treatmentrdquo Journal of Ophthal-mology vol 2010 Article ID 608751 8 pages 2010

[3] I G Obrosova S S M Chung and P F Kador ldquoDiabeticcataracts mechanisms and managementrdquo DiabetesMetabolismResearch and Reviews vol 26 no 3 pp 172ndash180 2010

[4] J Ding and T Y Wong ldquoCurrent epidemiology of diabeticretinopathy and diabetic macular edemardquo Current DiabetesReports vol 12 no 4 pp 346ndash354 2012

[5] A Y W Lee and S S M Chung ldquoContributions of polyolpathway to oxidative stress in diabetic cataractrdquo The FASEBJournal vol 13 no 1 pp 23ndash30 1999

[6] S A Madsen-Bouterse and R A Kowluru ldquoOxidative stressand diabetic retinopathy pathophysiological mechanisms andtreatment perspectivesrdquo Reviews in Endocrine and MetabolicDisorders vol 9 no 4 pp 315ndash327 2008

[7] I G Obrosova and P F Kador ldquoAldose reductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 1389ndash2010 2011

[8] V R Drel P Pacher T K Ali et al ldquoAldose reductase inhibitorfidarestat counteracts diabetes-associated cataract formationretinal oxidative-nitrosative stress glial activation and apopto-sisrdquo International Journal of Molecular Medicine vol 21 no 6pp 667ndash676 2008

[9] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (Purple Waxy Corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[10] Y Morimitsu K Kubota T Tashiro E Hashizume T Kamiyaand T Osawa ldquoInhibitory effect of anthocyanins and coloredrice ondiabetic cataract formation in the rat lensesrdquo Interna-tional Congress Series vol 1245 pp 503ndash508 2002

[11] M Saraswat P Suryanarayana P Y Reddy M A Patil NBalakrishna and G B Reddy ldquoAntiglycating potential of Zin-giber officinalis and delay of diabetic cataract in ratsrdquoMolecularVision vol 16 pp 1525ndash1537 2010

[12] Y Li V H Tran C C Duke and B D Roufogalis ldquoPreventiveand protective properties of Zingiber officinale (Ginger) indiabetes mellitus diabetic complications and associated lipidand other metabolic disorders a brief reviewrdquo Evidence-basedComplementary and Alternative Medicine vol 2012 Article ID516870 10 pages 2012

[13] P Suryanarayana M Saraswat T Mrudula T P Krishna KKrishnaswamy andG B Reddy ldquoCurcumin and turmeric delaystreptozotocin-induced diabetic cataract in ratsrdquo InvestigativeOphthalmology amp Visual Science vol 46 no 6 pp 2092ndash20992005

[14] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[15] Y Sun L W Oberley and Y Li ldquoA simple method for clinicalassay of superoxide dismutaserdquo Clinical Chemistry vol 34 no3 pp 497ndash500 1988

[16] L Goth ldquoA simple method for determination of serum catalaseactivity and revision of reference rangerdquo Clinica Chimica Actavol 196 no 2-3 pp 143ndash151 1991

[17] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973

[18] M B Patel and S M Mishra ldquoAldose reductase inhibitoryactivity and anti catraract potential of some traditionallyacclaimed antidiabetic medicinal plantsrdquo Oriental Pharmacyand Experimental Medicine vol 9 no 3 pp 245ndash251 2009

[19] R Agarwal I Iezhitsa N A Awaludin et al ldquoEffects ofmagnesium taurate on the onset and progression of galactose-induced experimental cataract in vivo and in vitro evaluationrdquoExperimental Eye Research vol 110 pp 35ndash43 2013

[20] P M Martin P Roon T K van Ells V Ganapathy and SB Smith ldquoDeath of retinal neurons in streptozotocin-induceddiabetic micerdquo Investigative Ophthalmology and Visual Sciencevol 45 no 9 pp 3330ndash3336 2004

[21] H W van Dijk F D Verbraak P H B Kok et al ldquoDecreasedretinal ganglion cell layer thickness in patients with type 1diabetesrdquo Investigative Ophthalmology and Visual Science vol51 no 7 pp 3660ndash3665 2010

[22] T S Kern and A J Barber ldquoRetinal ganglion cells in diabetesrdquoJournal of Physiology vol 586 no 18 pp 4401ndash4408 2008

[23] G A Ramadan ldquoSorbitol-induced diabetic-like retinal lesionsin rats microscopic studyrdquo American Journal of Pharmacologyand Toxicology vol 2 no 2 pp 89ndash97 2007

[24] B Ozmen D Ozmen E Erkin I Guner S Habif and OBaymdir ldquoLens superoxide dismutase and catalase activities indiabetic cataractrdquoClinical Biochemistry vol 35 no 1 pp 69ndash722002

[25] A B M Reddy R Tammali R Mishra S Srivastava S KSrivastava and K V Ramana ldquoAldose reductase deficiencyprotects sugar-induced lens opacification in ratsrdquo Chemico-Biological Interactions vol 191 no 1ndash3 pp 346ndash350 2011

[26] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (purple waxy corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[27] I G Obrosova and P F Kador ldquoAldosereductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 373ndash385 2011

[28] PMaher andAHanneken ldquoFlavonoids protect retinal ganglioncells from oxidative stress-induced deathrdquo Investigative Oph-thalmology and Visual Science vol 46 no 12 pp 4796ndash48032005

[29] M Stefek ldquoNatural flavonoids as potential multifunctionalagents in prevention of diabetic cataractrdquo InterdisciplinaryToxicology vol 4 no 2 pp 69ndash77 2011

Oxidative Medicine and Cellular Longevity 11

[30] S F Nabavi S Habtemariam M Daglia N Shafighi A JBarber and S M Nabavi ldquoAnthocyanins as a potential therapyfor diabetic retinopathyrdquo Current Medicinal Chemistry vol 22no 1 pp 51ndash58 2014

[31] A H Rahmani F M Al Shabrmi and S M Aly ldquoActiveingredients of ginger as potential candidates in the preventionand treatment of diseases via modulation of biological activ-itiesrdquo International Journal of Physiology Pathophysiology andPharmacology vol 6 no 2 pp 125ndash136 2014

[32] H Watanabe H Kosano and H Nishigori ldquoSteroid-inducedshort term diabetes in chick embryo reversible effects of insulinon metabolic changes and cataract formationrdquo InvestigativeOphthalmology amp Visual Science vol 41 no 7 pp 1846ndash18522000

[33] E R James ldquoThe etiology of steroid cataractrdquo Journal of OcularPharmacology and Therapeutics vol 23 no 5 pp 403ndash4202007

[34] P Greaves Histopathological of Preclinical Toxicity StudiesElsevier New York NY USA 3rd edition 2007

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

8 Oxidative Medicine and Cellular Longevity

0002000400060008000

100001200014000160001800020000

lowastlowastlowastlowastlowastlowast

lowastlowastlowast

Tota

l ret

inal

thic

knes

s (120583

m)

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 6 The total retinal thickness of retina (TRT) of control anddiabetic rats (DM) which received either vehicle or the combinationextract of purple waxy corn and ginger (PWCG) at doses of 50 100and 200mgsdotkgminus1 BW for 10weeks 119875 valuelt 0001 comparedwithcontrol and lowastlowastlowast119875 value lt 0001 compared with vehicle

000500

100015002000250030003500400045005000

The r

etin

al n

ucle

ar la

yer t

hick

ness

(120583m

)

lowastlowastlowast lowastlowastlowastlowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 7 The thickness of the retinal outer nuclear layer (ROT)of control and diabetic rats (DM) which received either vehicle orthe combination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks

119875 valuelt 0001 compared with control and lowastlowastlowast119875 value lt 0001 comparedwith vehicle

during the slit lamp examination in humanThismay possiblyinfluence the precision of grading scores which reflect thereal severity of cataract To obtain the precise severity ofcataract the slit lamp examination should be performedwhile pupil was dilated However all rats were assessed underthe condition without the pupil dilator so no confoundingerror about the beneficial effect of PWCG on cataract wasachieved

The lenticular opacification in control rats was alsoobserved during the 6ndash10 weeks of study period Based onthe previous study that stress hormone or glucocorticoids

000500

100015002000250030003500400045005000

The n

umbe

r of n

euro

nin

gan

glio

n ce

ll lay

er in

retin

a

lowastlowastlowastlowastlowastlowast

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 8 The number of cells in the ganglion cell layer (NG)in retina of control and diabetic rats (DM) which received eithervehicle or the combination extract of purple waxy corn and ginger(PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks 119875value lt 0001 compared with control and lowast lowastlowastlowast119875 value lt 005 and0001 compared with vehicle

MD

A (U

mg

prot

ein)

000

100

200

300

400

500

600

lowastlowastlowast lowastlowastlowastlowastlowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 9 The level of malondialdehyde (MDA) (Umg protein) inlens of control and diabetic rats (DM) which received either vehicleor the combination extract of purple waxy corn and ginger (PWCG)at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 = 8group)119875 value lt 0001 compared with control and lowastlowastlowast119875 value lt 0001

compared with diabetic rats which received vehicle (DM + vehicle)

can also induce cataract [32 33] we do suggest that thedevelopment of cataract in control group may possibly occuras the exposure to stress condition leading to the elevationof glucocorticoids hormone resulting in the cataractogenesisUnfortunately the level of corticosterone has not been deter-mined in this study Therefore the precise understandingmechanism of this change requires further investigation Inaddition spontaneous age-related cataract can also occurin laboratory rodents The prevalence is varied dependingon species Even young adolescent rodents can be found tohave cataract approximately 20 [34]Therefore the cataract

Oxidative Medicine and Cellular Longevity 9SO

D (U

mg

prot

ein)

000050100150200250300350400450

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 10 The activity of superoxide dismutase (SOD) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BWfor 10weeks(119873 = 8group) 119875 value lt 0001 compared with control

CAT

(Um

g pr

otei

n)

000

1000

2000

3000

4000

5000

6000

lowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 11 The activity of catalase (CAT) (Umg protein) in lensof control and diabetic rats (DM) which received either vehicle orthe combination extract of purple waxy corn and ginger (PWCG) atdoses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 = 8group)119875 value lt 0001 compared with control and lowastlowast119875 value lt 001

compared with diabetic rats which received vehicle (DM + vehicle)

observed in normal rats might occur either via stress orvia spontaneous cataractogenesis of the laboratory rodentsmentioned earlier

5 Conclusion

The combination extract of purple waxy corn seeds andginger rhizome (PWCG) can protect against diabetic cataractand diabetic retinopathyThepossible underlyingmechanismmay occur partly via the decreased oxidative stress and thesuppression of aldose reductase activity in lens The possibleactive ingredient may be associated with flavonoids such as

000020040060080100120140160180200

GPx

(Um

g pr

otei

n)

lowastlowastlowast

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 12 The activity of glutathione peroxidase (GPx) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10weeks (119873 = 8group) 119875 value lt 0001 compared with controland lowast lowastlowastlowast119875 value lt 005 0001 respectively compared with diabeticrats which received vehicle (DM + vehicle)

AR

activ

ity (n

mol

min

mg)

00000

00010

00020

00030

00040

00050

00060

00070

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 13 The activity of aldose reductase (AR) (nmolminmg)in lens of control and diabetic rats (DM) which received eithervehicle or the combination extract of purple waxy corn and ginger(PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 =8group) 119875 value lt 005 compared with control and lowast119875 value lt005 compared with diabetic rats which received vehicle (DM +vehicle)

anthocyanin and other flavonoids Further researches are stillessential to understand the precise underlying mechanismand possible active ingredient

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

10 Oxidative Medicine and Cellular Longevity

Acknowledgments

This study was supported by the Higher Education ResearchPromotion andNational Research University Project ofThai-land Office of the Higher Education Commission throughthe Food and Functional Food Research Cluster of KhonKaen University and the Integrative Complementary Alter-native Medicine Research and Development Center KhonKaen University Thailand Great thanks were also extendedto Research and Development Center of Herbal HealthProduct for providing the opportunity to collaborate in thesubcluster

References

[1] R L Engerman and T S Kern ldquoHyperglycemia as a cause ofdiabetic retinopathyrdquo Metabolism vol 35 supplement 1 no 4pp 20ndash23 1986

[2] A Pollreisz and U Schmidt-Erfurth ldquoDiabetic cataractmdashpathogenesis epidemiology and treatmentrdquo Journal of Ophthal-mology vol 2010 Article ID 608751 8 pages 2010

[3] I G Obrosova S S M Chung and P F Kador ldquoDiabeticcataracts mechanisms and managementrdquo DiabetesMetabolismResearch and Reviews vol 26 no 3 pp 172ndash180 2010

[4] J Ding and T Y Wong ldquoCurrent epidemiology of diabeticretinopathy and diabetic macular edemardquo Current DiabetesReports vol 12 no 4 pp 346ndash354 2012

[5] A Y W Lee and S S M Chung ldquoContributions of polyolpathway to oxidative stress in diabetic cataractrdquo The FASEBJournal vol 13 no 1 pp 23ndash30 1999

[6] S A Madsen-Bouterse and R A Kowluru ldquoOxidative stressand diabetic retinopathy pathophysiological mechanisms andtreatment perspectivesrdquo Reviews in Endocrine and MetabolicDisorders vol 9 no 4 pp 315ndash327 2008

[7] I G Obrosova and P F Kador ldquoAldose reductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 1389ndash2010 2011

[8] V R Drel P Pacher T K Ali et al ldquoAldose reductase inhibitorfidarestat counteracts diabetes-associated cataract formationretinal oxidative-nitrosative stress glial activation and apopto-sisrdquo International Journal of Molecular Medicine vol 21 no 6pp 667ndash676 2008

[9] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (Purple Waxy Corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[10] Y Morimitsu K Kubota T Tashiro E Hashizume T Kamiyaand T Osawa ldquoInhibitory effect of anthocyanins and coloredrice ondiabetic cataract formation in the rat lensesrdquo Interna-tional Congress Series vol 1245 pp 503ndash508 2002

[11] M Saraswat P Suryanarayana P Y Reddy M A Patil NBalakrishna and G B Reddy ldquoAntiglycating potential of Zin-giber officinalis and delay of diabetic cataract in ratsrdquoMolecularVision vol 16 pp 1525ndash1537 2010

[12] Y Li V H Tran C C Duke and B D Roufogalis ldquoPreventiveand protective properties of Zingiber officinale (Ginger) indiabetes mellitus diabetic complications and associated lipidand other metabolic disorders a brief reviewrdquo Evidence-basedComplementary and Alternative Medicine vol 2012 Article ID516870 10 pages 2012

[13] P Suryanarayana M Saraswat T Mrudula T P Krishna KKrishnaswamy andG B Reddy ldquoCurcumin and turmeric delaystreptozotocin-induced diabetic cataract in ratsrdquo InvestigativeOphthalmology amp Visual Science vol 46 no 6 pp 2092ndash20992005

[14] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[15] Y Sun L W Oberley and Y Li ldquoA simple method for clinicalassay of superoxide dismutaserdquo Clinical Chemistry vol 34 no3 pp 497ndash500 1988

[16] L Goth ldquoA simple method for determination of serum catalaseactivity and revision of reference rangerdquo Clinica Chimica Actavol 196 no 2-3 pp 143ndash151 1991

[17] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973

[18] M B Patel and S M Mishra ldquoAldose reductase inhibitoryactivity and anti catraract potential of some traditionallyacclaimed antidiabetic medicinal plantsrdquo Oriental Pharmacyand Experimental Medicine vol 9 no 3 pp 245ndash251 2009

[19] R Agarwal I Iezhitsa N A Awaludin et al ldquoEffects ofmagnesium taurate on the onset and progression of galactose-induced experimental cataract in vivo and in vitro evaluationrdquoExperimental Eye Research vol 110 pp 35ndash43 2013

[20] P M Martin P Roon T K van Ells V Ganapathy and SB Smith ldquoDeath of retinal neurons in streptozotocin-induceddiabetic micerdquo Investigative Ophthalmology and Visual Sciencevol 45 no 9 pp 3330ndash3336 2004

[21] H W van Dijk F D Verbraak P H B Kok et al ldquoDecreasedretinal ganglion cell layer thickness in patients with type 1diabetesrdquo Investigative Ophthalmology and Visual Science vol51 no 7 pp 3660ndash3665 2010

[22] T S Kern and A J Barber ldquoRetinal ganglion cells in diabetesrdquoJournal of Physiology vol 586 no 18 pp 4401ndash4408 2008

[23] G A Ramadan ldquoSorbitol-induced diabetic-like retinal lesionsin rats microscopic studyrdquo American Journal of Pharmacologyand Toxicology vol 2 no 2 pp 89ndash97 2007

[24] B Ozmen D Ozmen E Erkin I Guner S Habif and OBaymdir ldquoLens superoxide dismutase and catalase activities indiabetic cataractrdquoClinical Biochemistry vol 35 no 1 pp 69ndash722002

[25] A B M Reddy R Tammali R Mishra S Srivastava S KSrivastava and K V Ramana ldquoAldose reductase deficiencyprotects sugar-induced lens opacification in ratsrdquo Chemico-Biological Interactions vol 191 no 1ndash3 pp 346ndash350 2011

[26] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (purple waxy corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[27] I G Obrosova and P F Kador ldquoAldosereductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 373ndash385 2011

[28] PMaher andAHanneken ldquoFlavonoids protect retinal ganglioncells from oxidative stress-induced deathrdquo Investigative Oph-thalmology and Visual Science vol 46 no 12 pp 4796ndash48032005

[29] M Stefek ldquoNatural flavonoids as potential multifunctionalagents in prevention of diabetic cataractrdquo InterdisciplinaryToxicology vol 4 no 2 pp 69ndash77 2011

Oxidative Medicine and Cellular Longevity 11

[30] S F Nabavi S Habtemariam M Daglia N Shafighi A JBarber and S M Nabavi ldquoAnthocyanins as a potential therapyfor diabetic retinopathyrdquo Current Medicinal Chemistry vol 22no 1 pp 51ndash58 2014

[31] A H Rahmani F M Al Shabrmi and S M Aly ldquoActiveingredients of ginger as potential candidates in the preventionand treatment of diseases via modulation of biological activ-itiesrdquo International Journal of Physiology Pathophysiology andPharmacology vol 6 no 2 pp 125ndash136 2014

[32] H Watanabe H Kosano and H Nishigori ldquoSteroid-inducedshort term diabetes in chick embryo reversible effects of insulinon metabolic changes and cataract formationrdquo InvestigativeOphthalmology amp Visual Science vol 41 no 7 pp 1846ndash18522000

[33] E R James ldquoThe etiology of steroid cataractrdquo Journal of OcularPharmacology and Therapeutics vol 23 no 5 pp 403ndash4202007

[34] P Greaves Histopathological of Preclinical Toxicity StudiesElsevier New York NY USA 3rd edition 2007

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Oxidative Medicine and Cellular Longevity 9SO

D (U

mg

prot

ein)

000050100150200250300350400450

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 10 The activity of superoxide dismutase (SOD) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BWfor 10weeks(119873 = 8group) 119875 value lt 0001 compared with control

CAT

(Um

g pr

otei

n)

000

1000

2000

3000

4000

5000

6000

lowastlowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 11 The activity of catalase (CAT) (Umg protein) in lensof control and diabetic rats (DM) which received either vehicle orthe combination extract of purple waxy corn and ginger (PWCG) atdoses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 = 8group)119875 value lt 0001 compared with control and lowastlowast119875 value lt 001

compared with diabetic rats which received vehicle (DM + vehicle)

observed in normal rats might occur either via stress orvia spontaneous cataractogenesis of the laboratory rodentsmentioned earlier

5 Conclusion

The combination extract of purple waxy corn seeds andginger rhizome (PWCG) can protect against diabetic cataractand diabetic retinopathyThepossible underlyingmechanismmay occur partly via the decreased oxidative stress and thesuppression of aldose reductase activity in lens The possibleactive ingredient may be associated with flavonoids such as

000020040060080100120140160180200

GPx

(Um

g pr

otei

n)

lowastlowastlowast

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 12 The activity of glutathione peroxidase (GPx) (Umgprotein) in lens of control and diabetic rats (DM) which receivedeither vehicle or the combination extract of purple waxy corn andginger (PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10weeks (119873 = 8group) 119875 value lt 0001 compared with controland lowast lowastlowastlowast119875 value lt 005 0001 respectively compared with diabeticrats which received vehicle (DM + vehicle)

AR

activ

ity (n

mol

min

mg)

00000

00010

00020

00030

00040

00050

00060

00070

lowast

ControlDM + vehicle

DM + PWCG (100

DM + PWCG (200

DM + PWCG (50 mgkgmiddotBWminus1)mgkgmiddotBWminus1)mgkgmiddotBWminus1)

Figure 13 The activity of aldose reductase (AR) (nmolminmg)in lens of control and diabetic rats (DM) which received eithervehicle or the combination extract of purple waxy corn and ginger(PWCG) at doses of 50 100 and 200mgsdotkgminus1 BW for 10 weeks (119873 =8group) 119875 value lt 005 compared with control and lowast119875 value lt005 compared with diabetic rats which received vehicle (DM +vehicle)

anthocyanin and other flavonoids Further researches are stillessential to understand the precise underlying mechanismand possible active ingredient

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

10 Oxidative Medicine and Cellular Longevity

Acknowledgments

This study was supported by the Higher Education ResearchPromotion andNational Research University Project ofThai-land Office of the Higher Education Commission throughthe Food and Functional Food Research Cluster of KhonKaen University and the Integrative Complementary Alter-native Medicine Research and Development Center KhonKaen University Thailand Great thanks were also extendedto Research and Development Center of Herbal HealthProduct for providing the opportunity to collaborate in thesubcluster

References

[1] R L Engerman and T S Kern ldquoHyperglycemia as a cause ofdiabetic retinopathyrdquo Metabolism vol 35 supplement 1 no 4pp 20ndash23 1986

[2] A Pollreisz and U Schmidt-Erfurth ldquoDiabetic cataractmdashpathogenesis epidemiology and treatmentrdquo Journal of Ophthal-mology vol 2010 Article ID 608751 8 pages 2010

[3] I G Obrosova S S M Chung and P F Kador ldquoDiabeticcataracts mechanisms and managementrdquo DiabetesMetabolismResearch and Reviews vol 26 no 3 pp 172ndash180 2010

[4] J Ding and T Y Wong ldquoCurrent epidemiology of diabeticretinopathy and diabetic macular edemardquo Current DiabetesReports vol 12 no 4 pp 346ndash354 2012

[5] A Y W Lee and S S M Chung ldquoContributions of polyolpathway to oxidative stress in diabetic cataractrdquo The FASEBJournal vol 13 no 1 pp 23ndash30 1999

[6] S A Madsen-Bouterse and R A Kowluru ldquoOxidative stressand diabetic retinopathy pathophysiological mechanisms andtreatment perspectivesrdquo Reviews in Endocrine and MetabolicDisorders vol 9 no 4 pp 315ndash327 2008

[7] I G Obrosova and P F Kador ldquoAldose reductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 1389ndash2010 2011

[8] V R Drel P Pacher T K Ali et al ldquoAldose reductase inhibitorfidarestat counteracts diabetes-associated cataract formationretinal oxidative-nitrosative stress glial activation and apopto-sisrdquo International Journal of Molecular Medicine vol 21 no 6pp 667ndash676 2008

[9] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (Purple Waxy Corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[10] Y Morimitsu K Kubota T Tashiro E Hashizume T Kamiyaand T Osawa ldquoInhibitory effect of anthocyanins and coloredrice ondiabetic cataract formation in the rat lensesrdquo Interna-tional Congress Series vol 1245 pp 503ndash508 2002

[11] M Saraswat P Suryanarayana P Y Reddy M A Patil NBalakrishna and G B Reddy ldquoAntiglycating potential of Zin-giber officinalis and delay of diabetic cataract in ratsrdquoMolecularVision vol 16 pp 1525ndash1537 2010

[12] Y Li V H Tran C C Duke and B D Roufogalis ldquoPreventiveand protective properties of Zingiber officinale (Ginger) indiabetes mellitus diabetic complications and associated lipidand other metabolic disorders a brief reviewrdquo Evidence-basedComplementary and Alternative Medicine vol 2012 Article ID516870 10 pages 2012

[13] P Suryanarayana M Saraswat T Mrudula T P Krishna KKrishnaswamy andG B Reddy ldquoCurcumin and turmeric delaystreptozotocin-induced diabetic cataract in ratsrdquo InvestigativeOphthalmology amp Visual Science vol 46 no 6 pp 2092ndash20992005

[14] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[15] Y Sun L W Oberley and Y Li ldquoA simple method for clinicalassay of superoxide dismutaserdquo Clinical Chemistry vol 34 no3 pp 497ndash500 1988

[16] L Goth ldquoA simple method for determination of serum catalaseactivity and revision of reference rangerdquo Clinica Chimica Actavol 196 no 2-3 pp 143ndash151 1991

[17] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973

[18] M B Patel and S M Mishra ldquoAldose reductase inhibitoryactivity and anti catraract potential of some traditionallyacclaimed antidiabetic medicinal plantsrdquo Oriental Pharmacyand Experimental Medicine vol 9 no 3 pp 245ndash251 2009

[19] R Agarwal I Iezhitsa N A Awaludin et al ldquoEffects ofmagnesium taurate on the onset and progression of galactose-induced experimental cataract in vivo and in vitro evaluationrdquoExperimental Eye Research vol 110 pp 35ndash43 2013

[20] P M Martin P Roon T K van Ells V Ganapathy and SB Smith ldquoDeath of retinal neurons in streptozotocin-induceddiabetic micerdquo Investigative Ophthalmology and Visual Sciencevol 45 no 9 pp 3330ndash3336 2004

[21] H W van Dijk F D Verbraak P H B Kok et al ldquoDecreasedretinal ganglion cell layer thickness in patients with type 1diabetesrdquo Investigative Ophthalmology and Visual Science vol51 no 7 pp 3660ndash3665 2010

[22] T S Kern and A J Barber ldquoRetinal ganglion cells in diabetesrdquoJournal of Physiology vol 586 no 18 pp 4401ndash4408 2008

[23] G A Ramadan ldquoSorbitol-induced diabetic-like retinal lesionsin rats microscopic studyrdquo American Journal of Pharmacologyand Toxicology vol 2 no 2 pp 89ndash97 2007

[24] B Ozmen D Ozmen E Erkin I Guner S Habif and OBaymdir ldquoLens superoxide dismutase and catalase activities indiabetic cataractrdquoClinical Biochemistry vol 35 no 1 pp 69ndash722002

[25] A B M Reddy R Tammali R Mishra S Srivastava S KSrivastava and K V Ramana ldquoAldose reductase deficiencyprotects sugar-induced lens opacification in ratsrdquo Chemico-Biological Interactions vol 191 no 1ndash3 pp 346ndash350 2011

[26] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (purple waxy corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[27] I G Obrosova and P F Kador ldquoAldosereductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 373ndash385 2011

[28] PMaher andAHanneken ldquoFlavonoids protect retinal ganglioncells from oxidative stress-induced deathrdquo Investigative Oph-thalmology and Visual Science vol 46 no 12 pp 4796ndash48032005

[29] M Stefek ldquoNatural flavonoids as potential multifunctionalagents in prevention of diabetic cataractrdquo InterdisciplinaryToxicology vol 4 no 2 pp 69ndash77 2011

Oxidative Medicine and Cellular Longevity 11

[30] S F Nabavi S Habtemariam M Daglia N Shafighi A JBarber and S M Nabavi ldquoAnthocyanins as a potential therapyfor diabetic retinopathyrdquo Current Medicinal Chemistry vol 22no 1 pp 51ndash58 2014

[31] A H Rahmani F M Al Shabrmi and S M Aly ldquoActiveingredients of ginger as potential candidates in the preventionand treatment of diseases via modulation of biological activ-itiesrdquo International Journal of Physiology Pathophysiology andPharmacology vol 6 no 2 pp 125ndash136 2014

[32] H Watanabe H Kosano and H Nishigori ldquoSteroid-inducedshort term diabetes in chick embryo reversible effects of insulinon metabolic changes and cataract formationrdquo InvestigativeOphthalmology amp Visual Science vol 41 no 7 pp 1846ndash18522000

[33] E R James ldquoThe etiology of steroid cataractrdquo Journal of OcularPharmacology and Therapeutics vol 23 no 5 pp 403ndash4202007

[34] P Greaves Histopathological of Preclinical Toxicity StudiesElsevier New York NY USA 3rd edition 2007

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

10 Oxidative Medicine and Cellular Longevity

Acknowledgments

This study was supported by the Higher Education ResearchPromotion andNational Research University Project ofThai-land Office of the Higher Education Commission throughthe Food and Functional Food Research Cluster of KhonKaen University and the Integrative Complementary Alter-native Medicine Research and Development Center KhonKaen University Thailand Great thanks were also extendedto Research and Development Center of Herbal HealthProduct for providing the opportunity to collaborate in thesubcluster

References

[1] R L Engerman and T S Kern ldquoHyperglycemia as a cause ofdiabetic retinopathyrdquo Metabolism vol 35 supplement 1 no 4pp 20ndash23 1986

[2] A Pollreisz and U Schmidt-Erfurth ldquoDiabetic cataractmdashpathogenesis epidemiology and treatmentrdquo Journal of Ophthal-mology vol 2010 Article ID 608751 8 pages 2010

[3] I G Obrosova S S M Chung and P F Kador ldquoDiabeticcataracts mechanisms and managementrdquo DiabetesMetabolismResearch and Reviews vol 26 no 3 pp 172ndash180 2010

[4] J Ding and T Y Wong ldquoCurrent epidemiology of diabeticretinopathy and diabetic macular edemardquo Current DiabetesReports vol 12 no 4 pp 346ndash354 2012

[5] A Y W Lee and S S M Chung ldquoContributions of polyolpathway to oxidative stress in diabetic cataractrdquo The FASEBJournal vol 13 no 1 pp 23ndash30 1999

[6] S A Madsen-Bouterse and R A Kowluru ldquoOxidative stressand diabetic retinopathy pathophysiological mechanisms andtreatment perspectivesrdquo Reviews in Endocrine and MetabolicDisorders vol 9 no 4 pp 315ndash327 2008

[7] I G Obrosova and P F Kador ldquoAldose reductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 1389ndash2010 2011

[8] V R Drel P Pacher T K Ali et al ldquoAldose reductase inhibitorfidarestat counteracts diabetes-associated cataract formationretinal oxidative-nitrosative stress glial activation and apopto-sisrdquo International Journal of Molecular Medicine vol 21 no 6pp 667ndash676 2008

[9] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (Purple Waxy Corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[10] Y Morimitsu K Kubota T Tashiro E Hashizume T Kamiyaand T Osawa ldquoInhibitory effect of anthocyanins and coloredrice ondiabetic cataract formation in the rat lensesrdquo Interna-tional Congress Series vol 1245 pp 503ndash508 2002

[11] M Saraswat P Suryanarayana P Y Reddy M A Patil NBalakrishna and G B Reddy ldquoAntiglycating potential of Zin-giber officinalis and delay of diabetic cataract in ratsrdquoMolecularVision vol 16 pp 1525ndash1537 2010

[12] Y Li V H Tran C C Duke and B D Roufogalis ldquoPreventiveand protective properties of Zingiber officinale (Ginger) indiabetes mellitus diabetic complications and associated lipidand other metabolic disorders a brief reviewrdquo Evidence-basedComplementary and Alternative Medicine vol 2012 Article ID516870 10 pages 2012

[13] P Suryanarayana M Saraswat T Mrudula T P Krishna KKrishnaswamy andG B Reddy ldquoCurcumin and turmeric delaystreptozotocin-induced diabetic cataract in ratsrdquo InvestigativeOphthalmology amp Visual Science vol 46 no 6 pp 2092ndash20992005

[14] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[15] Y Sun L W Oberley and Y Li ldquoA simple method for clinicalassay of superoxide dismutaserdquo Clinical Chemistry vol 34 no3 pp 497ndash500 1988

[16] L Goth ldquoA simple method for determination of serum catalaseactivity and revision of reference rangerdquo Clinica Chimica Actavol 196 no 2-3 pp 143ndash151 1991

[17] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973

[18] M B Patel and S M Mishra ldquoAldose reductase inhibitoryactivity and anti catraract potential of some traditionallyacclaimed antidiabetic medicinal plantsrdquo Oriental Pharmacyand Experimental Medicine vol 9 no 3 pp 245ndash251 2009

[19] R Agarwal I Iezhitsa N A Awaludin et al ldquoEffects ofmagnesium taurate on the onset and progression of galactose-induced experimental cataract in vivo and in vitro evaluationrdquoExperimental Eye Research vol 110 pp 35ndash43 2013

[20] P M Martin P Roon T K van Ells V Ganapathy and SB Smith ldquoDeath of retinal neurons in streptozotocin-induceddiabetic micerdquo Investigative Ophthalmology and Visual Sciencevol 45 no 9 pp 3330ndash3336 2004

[21] H W van Dijk F D Verbraak P H B Kok et al ldquoDecreasedretinal ganglion cell layer thickness in patients with type 1diabetesrdquo Investigative Ophthalmology and Visual Science vol51 no 7 pp 3660ndash3665 2010

[22] T S Kern and A J Barber ldquoRetinal ganglion cells in diabetesrdquoJournal of Physiology vol 586 no 18 pp 4401ndash4408 2008

[23] G A Ramadan ldquoSorbitol-induced diabetic-like retinal lesionsin rats microscopic studyrdquo American Journal of Pharmacologyand Toxicology vol 2 no 2 pp 89ndash97 2007

[24] B Ozmen D Ozmen E Erkin I Guner S Habif and OBaymdir ldquoLens superoxide dismutase and catalase activities indiabetic cataractrdquoClinical Biochemistry vol 35 no 1 pp 69ndash722002

[25] A B M Reddy R Tammali R Mishra S Srivastava S KSrivastava and K V Ramana ldquoAldose reductase deficiencyprotects sugar-induced lens opacification in ratsrdquo Chemico-Biological Interactions vol 191 no 1ndash3 pp 346ndash350 2011

[26] P Thiraphatthanavong J Wattanathorn S Muchimapura etal ldquoPreventive effect of Zea mays L (purple waxy corn) onexperimental diabetic cataractrdquo BioMed Research Internationalvol 2014 Article ID 507435 8 pages 2014

[27] I G Obrosova and P F Kador ldquoAldosereductasepolyolinhibitors for diabetic retinopathyrdquo Current PharmaceuticalBiotechnology vol 12 no 3 pp 373ndash385 2011

[28] PMaher andAHanneken ldquoFlavonoids protect retinal ganglioncells from oxidative stress-induced deathrdquo Investigative Oph-thalmology and Visual Science vol 46 no 12 pp 4796ndash48032005

[29] M Stefek ldquoNatural flavonoids as potential multifunctionalagents in prevention of diabetic cataractrdquo InterdisciplinaryToxicology vol 4 no 2 pp 69ndash77 2011

Oxidative Medicine and Cellular Longevity 11

[30] S F Nabavi S Habtemariam M Daglia N Shafighi A JBarber and S M Nabavi ldquoAnthocyanins as a potential therapyfor diabetic retinopathyrdquo Current Medicinal Chemistry vol 22no 1 pp 51ndash58 2014

[31] A H Rahmani F M Al Shabrmi and S M Aly ldquoActiveingredients of ginger as potential candidates in the preventionand treatment of diseases via modulation of biological activ-itiesrdquo International Journal of Physiology Pathophysiology andPharmacology vol 6 no 2 pp 125ndash136 2014

[32] H Watanabe H Kosano and H Nishigori ldquoSteroid-inducedshort term diabetes in chick embryo reversible effects of insulinon metabolic changes and cataract formationrdquo InvestigativeOphthalmology amp Visual Science vol 41 no 7 pp 1846ndash18522000

[33] E R James ldquoThe etiology of steroid cataractrdquo Journal of OcularPharmacology and Therapeutics vol 23 no 5 pp 403ndash4202007

[34] P Greaves Histopathological of Preclinical Toxicity StudiesElsevier New York NY USA 3rd edition 2007

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Oxidative Medicine and Cellular Longevity 11

[30] S F Nabavi S Habtemariam M Daglia N Shafighi A JBarber and S M Nabavi ldquoAnthocyanins as a potential therapyfor diabetic retinopathyrdquo Current Medicinal Chemistry vol 22no 1 pp 51ndash58 2014

[31] A H Rahmani F M Al Shabrmi and S M Aly ldquoActiveingredients of ginger as potential candidates in the preventionand treatment of diseases via modulation of biological activ-itiesrdquo International Journal of Physiology Pathophysiology andPharmacology vol 6 no 2 pp 125ndash136 2014

[32] H Watanabe H Kosano and H Nishigori ldquoSteroid-inducedshort term diabetes in chick embryo reversible effects of insulinon metabolic changes and cataract formationrdquo InvestigativeOphthalmology amp Visual Science vol 41 no 7 pp 1846ndash18522000

[33] E R James ldquoThe etiology of steroid cataractrdquo Journal of OcularPharmacology and Therapeutics vol 23 no 5 pp 403ndash4202007

[34] P Greaves Histopathological of Preclinical Toxicity StudiesElsevier New York NY USA 3rd edition 2007

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom