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Chapter 7 Bacopa Monnieri

Deptt. of Med. Elemn. & Toxicology 132 PhD Thesis

7.1. Introduction

The marked susceptibility of the kidney to chemical damage has been shown as a

function of several contributing factors including its anatomical proximity to blood

supply, role in metabolism and excretion of xenobiotics. Hence prevention of renal

damage is of great concern. Potassium bromate (KBrO3) is a chemical oxidizing mediator

found in drinking water as disinfection by product of surface water ozonation (Kurokawa

et al., 1982). Potassium bromate (KBrO3) is widely used as a food additive in the bread-

making process for the maturation of flour because of its oxidizing properties and as a

neutralizer in cold-wave hair lotions (Kurokawa et al., 1982). It is also used in food

products and in the production of fish paste and fermented beverages. It has been shown

to produce ROS (Sai et al., 1992), induce lipid peroxidation (Kurokawa, 1990),

genotoxicity (Jaloszynski et al., 2007) 8-hydroxyguanosine (8-OH-dG) modification in

kidney DNA (Kasai et al., 1987), diminished mitochondrial function and overexpression

of glycolytic enzymes (Ahlborn et al., 2009). Nephrotoxicity and neurotoxicity are the

primary responses to KBrO3 with death occurring from solutions containing as little as 12

g KBrO3. Although there is no strong evidence available to support the potential

carcinogenicity of KBrO3 to humans, it is classified as a category 2B carcinogen

(possibly carcinogenic to humans) based on adequate data of kidney carcinogenicity in

rats by the International Agency for Research on Cancer (IARC). Most reports of human

toxicity include hearing defects and renal failure following unintentional or purposeful

ingestion of few grams of potassium bromate. (De Angelo et al., 1998) Chronic

exposures to potassium bromate at drinking water levels above 50 ppm cause renal cell

carcinomas in rats, hamsters and mice, and cause thyroid and mesothelioma tumors in

rats. It has been demonstrated that KBrO3 induces renal oxidative stress, renal cell

tumors, mesotheliomas of the peritoneum, and follicular cell tumors of the thyroid.

KBrO3- has also been reported to mediate generation of oxidative stress (McDorman et

al., 2005) which directs augmentation in renal cellular proliferation (Umemura et al.,

2004). RCC is the sixth leading cause of cancer deaths in the United States. Scientists

reporting the mechanism of potassium bromate-induced toxicity have focused on

oxidative stress as a key event in the induction of these carcinomas (McDorman et al.,

2005).



Bacopa monnieri Linn. (Scrophulariaceae) also known as “Brahmi” is a creeping

glabrous, succulent herb, rooting at nodes, distributed throughout Nepal, china, Taiwan,

Sri Lanka and India in all plain districts, ascending to an altitude of 1,320 m. Its

medicinal efficiency is extensively reported in Indian traditional literature such as Carak

Samhita, Athar-Ved, Susrutu Samhita (Kishore & Singh, 2005) for treatment of epilepsy,

insomnia, anxiety and as a mild sedative and memory enhancer (Ernst, 2006; Tripathi et

al., 1996). As an ayurvedic medicine it is used as a nerve tonic. It is an important

component of south Asian cuisine. It is thought to improve memory, intelligence and

performance of sense organs, and it has also been used to treat asthma (Das et al., 2002).

Besides, B. monnieri (Brahmi) displays antioxidant (Tripathi et al 1996 ; Rajani et al.,

2004) anti-inflammatory, (Channa et al., 2006), antihyperglycemic (Ghosh et al., 2008),

antiamnesic (Prabhakar et al., 2008), antistress (Chowdhuri et al., 2002), and anxiolytic

(Shanker, & Singh 2000) activities too in animals. Bacopa extract contains two prominent

constituents namely, bacoside-A (Singh et al., 1988) and bacoside-B (Basu et al., 1967;

Singh et al., 1988). Herpestien and flavonoids and brahmine are also reported in B.

monnieri (Hou et al., 2002).Bacopa monnieri has been found to be effective against

Nitrobenzene induced liver damage (Menon et al., 2010) and carcinogen induced

hepatotoxicity (Janani et al., 2009). Therefore, we investigated the prophylactic effect of

B. monnieri extract in rats subjected to KBrO3-induced oxidative stress, inflammation,

apoptosis and subsequent renal tumor promotion.

7.2. Results

7.2.1. In vitro results of Bacopa monnieri

Total polyphenolic content of B.monnieri extract was found to be 196 + 0.81 mg GAE/g

(Gallic acid equivalents/g) dry weight. For the measurements of the reducing ability, we

investigated the Fe3+to Fe2+ transformation in the presence of the different concentrations

of extracts using the method of Oyaizu (Oyaizu, 1986). The reducing capacity of a

compound may serve as a significant indicator of its potential antioxidant. In our present

study Reducing potential of B.monnieri increases with increase in absorbance when

compared to control. The addition of Fe2+ or Fe3+ is a common approach to



experimentally induce oxidative stress in vitro and has also been used in two of our

assays (induction of lipid peroxidation and DNA sugar damage). Data obtained from both

the experiments reveal that there is dose dependent inhibition of Lipid peroxidation and

DNA sugar damage by B.monnieri. DPPH scavenging activity is a specific parameter to

determine the antioxidant activity of plant extracts. B. Monnieri exhibits a significant

concentration dependent DPPH scavenging activity at 50, 100, 200, 500, 100 µg/ml

respectively (Fig 1) Increase in DPPH scavenging activity indicates better antioxidant

property of the plant material.

7.2.2. Effect of pretreatment of Bacopa monnieri extract on glutathione and

glutathione dependent enzymes

Table 1 shows the effect of B. monnieri on level of glutathione and glutathione

dependent enzymes in the renal tissue of nephrotoxic rats. The decreased activities of

GSH, GPX, GR and GST in the tissue of nephrotoxic rats significantly increased upon

exposure with B. monnieri (P<0.001). Among the two different doses (100 and 200

mg/kg BW) of B. monnieri, 200 mg/kg dose was more effective.

7.2.3. Effect of Bacopa monnieri on KBrO3 induced modulation in Catalase, MDA,

SOD and XO

The changes in the levels of lipid peroxidation products, SOD, catalase and XO in control

and experiment animals are depicted in Table 2. The levels of TBARS and lipid

hydroperoxides were significantly increased (p < 0.001). On the other hand, the levels of

catalase and XO were also significantly increased (p < 0.05) in KBrO3 treated rats.

Administration of B .monnieri higher dose only significantly decreased the levels of

these parameters (p < 0.001), to normal in kidney. SOD showed marked depletion on

KBrO3 exposure (P<0.001) these levels were restored to normal by B. monnieri treatment

at both the doses (P<0.001). Level of catalase was also estimated in all the treatment

groups and B. monnieri was showing improvement (P<0.001) in the level of catalase as

compared to toxicant treated group.



7.2.4. Pre-treatment of Bacopa monnieri restores the level of toxicity markers like

creatinine, BUN, LDH and GGT

Table 3 depicts the effect of BM on the level of toxicity markers. Treatment of KBrO3

induced significant increase in toxicity markers (p<0.001), both the doses of B. monnieri

were instrumental in significantly bringing back level of all the markers to normal.

7.2.5. Effect on inflammatory markers: COX-2, NO, MPO and TNF-α.

Table 4 represents the significant increase of nitrite (P<0.001) in KBrO3-treated group

and attenuation of nitrite levels following oral feeding of B. monnieri (P<0.001) in both

the treatment groups. The decrease was statistically significant compared to the values

observed in the KBrO3-treated group.MPO activity was also measured in renal tissue as a

marker of inflammatory responses in the kidney (Table 2). KBrO3 significantly increased

MPO activity (P<0.001) in renal tissue. BM supplementation significantly (P<0.01,

P<0.001) inhibited the KBrO3-induced increase in renal MPO activity (Neutrophil

infiltration). COX-2 was barely detected in renal tissues of control rats. Renal sections

showed intense staining of COX-2 expression in KBrO3 exposed rats. Oral

supplementation of B. monnieri significantly ameliorated KBrO3-induced expression of

COX-2 (Figure 2).

7.2.6. Effect of BM extract on expression of NFĸB proteins in KBrO3-treated rats.

In case of control almost no expression of NFĸB protein was observed while KBrO3

treatment significantly enhanced the expression of NFĸB especially in the outer medulla

and inner cortical region. Pretreatment of B.monnieri at lower dose (100mg/kg BW) less

intense staining of medullary and corticular region while as higher dose of B.monnieri

(200mg/kg BW) showed almost complete suppression of this protein (Figure 5).

7.2.7. Effect of B.monnieri on level of Caspase-3 and p53 in the kidney of KBrO3

Treated rats

Instability of genome is basically linked to alterations in the control of apoptosis and to

changes in apoptosis-associated genes, including Caspase-3 and p53. Compared to the



Control group where there is minimal expression of p53, KBrO3-treatment increased the

expression of p53 (Figure 3). Consistent with data on oxidative stress, B.monnieri alone

did not change p53 expression. B.monnieri abolished the KBrO3-induced increase in p53

(Figure 3).Caspase-3 activity was also significantly elevated in the KBrO3-treated

(P<0.001) rat kidney. In the present study, we found that B.monnieri could markedly

decrease the activity of caspase-3 in KBrO3-treated rat kidney (P < 0.01) (Figure 6).The

pattern of change in Caspase-3 mimicked that of p53, as both proteins are engaged in

DNA-damage and repair processes.

7.2.8. Histopathological examination

Enhanced level of eosinophilic bodies characterized by large rounded droplets stained

strongly with eosin in the cytoplasm of proximal tubular epithelial cells is a well-known

KBrO3 treatment-related alteration in rat kidneys (Kurokawa et al., 1990). It is also

known to be associated with buildup of alpha-2u globulin and related to KBrO3-induced

cell proliferation and toxicities in rat kidneys. B. monnieri was successful in restoring the

deteriorated renal histomorphology to normal. (Figure 4)

7.2.9. The effect of pretreatment of animals with B.monnieri on KBrO3-mediated

induction of Tumor Promotion in terms of ODC activity and thymidine incorporation

Treatment with KBrO3 caused significant induction (P<0.001) in the ODC activity as

compared with saline-treated controls (Figure 7). The pretreatment of rats with

B.monnieri at a dose of 100 mg/kg body weight caused inhibition (P<0.05) in the

elevation of ODC activity, and at 200 mg/kg body weight caused more significant

(P<0.001) inhibition in the elevation of ODC activity as compared with KBrO3-treated

control group. Figure 8 also shows the effect of prophylaxis of rats with B.monnieri on

KBrO3 -mediated enhancement in the incorporation of [3H] thymidine into renal DNA.

Treatment with only KBrO3 caused significant increase (P<0.001) in the incorporation of

[3H] thymidine into renal DNA. B.monnieri pretreatment (100 and 200 mg/kg body

weight) caused significant reduction in the enhancement of DNA synthesis as compared

with KBrO3-treated group.



7.3. Discussion

In the present study, we investigated the efficacy of B.monnieri on KBrO3 induced renal

oxidative stress, inflammation, tumor promotion and apoptosis. B.monnieri ameliorates

oxidative stress, inflammation, and apoptosis induced by KBrO3 in renal tissue,Although

the mechanisms underlying the KBrO3-induced renal toxicity are not fully understood,

several investigators have shown that ROS are closely related to renal toxicity and

hyperproliferation induced by KBrO3 (Khan et al., 2004). Medicinal plants as component

of diet are used throughout the world for centuries to treat many diseases, and a major

portion of the world population relies on naturally occurring agents for their health

solutions. Plant-based antioxidants in food are not only efficient, but are also relatively

safe. Natural antioxidants have become increasingly essential as therapeutic agents

against oxidative stress and inflammatory disorders such as cancer (Khansari et al.,

2009). We have previously reported the role of oxidative stress in nephrotoxicity and

renal carcinogenesis by various toxicants like Fe-NTA, KBrO3, and NiCl2 (Laxmi et al.,

2006; Khan. et al., 2005; Jahangir, & Sultana, 2007; Iqbal, et al., 2009).

KBrO3 has been shown to increase the level of MDA. Lipid peroxidation and the

associated membrane damage is implicated in the pathophysiology of a number of

diseases, including renal carcinogenesis. Elevated lipid peroxidation has been reported

upon KBrO3 exposure as well (Sai et al., 1992).In the present study too KBrO3 treatment

enhanced the levels of MDA formation in renal tissue and B.monnieri significantly

ameliorated this increase. The observed prevention of MDA formation is in accordance

with previous reports were in B.monnieri has been demonstrated to inhibit MDA in liver

tissue. (Ghosh, et al., 2007)

The toxicity of KBrO3 is also due to its affinity for thiol (-SH) groups owing to which, it

can exhaust cellular GSH and cause damage to proteins and thiol enzymes. In addition,

GSH levels may also attenuate during neutralization of oxidative stress generated by

KBrO3. Under these conditions, B.monnieri extract may be expected to help in easing

oxidative stress burden, thus preserving the GSH levels. This may account, at least

partially, for the observed protection against KBrO3 induced decline in GSH levels.



Antioxidant enzymes and GSH play an important role in KBrO3 induced nephrotoxicity.

Watanabe et al., (2004) showed KBrO3 to decrease important antioxidant enzyme;

Glutathione Peroxidase which is in agreement with report by Sai, et al., (1992),

Umemura, et al., (1995) and many others stating that there was overall decrease in the

activity of all antioxidant enzymes further supporting that KBrO3 has direct inhibitory

effect on the endogenous physiological defense system (Farombi, et al., 2002).

B.monnieri supplementation restored the level of GSH and the activities of GST and

glutathione redox cycle enzymes in renal tissue. The activities of other antioxidant

enzymes like SOD, catalase and quinone reductase were also depleted by KBrO3

treatment. B.monnieri pretreatment significantly preserved the activities of all these

antioxidant enzymes. These findings were in a agreement with the report of Kapoor, et

al., (2009).

ROS could also induce damage to DNA, proteins and lipids within cells, which can lead

to tissue injury (Liu et al., 2010). KBrO3 being strong oxidizing agent has been shown to

cause oxidative modification of DNA bases, lipids and proteins in kidneys (Karbownik et

al. 2006). Cadenas, & Barja (1999) reported that KBrO3 increased the levels of 8-OHdG

in renal genomic DNA. These cells with modified DNA are either repaired by DNA

repair enzymes or are removed by apoptotic process. Apoptosis (programmed cell death)

is a key mechanism of cellular defense in sinking the risks of error-prone repair.

Increased ROS levels contribute to the apoptotic cell death every time they are generated

in the context of the apoptotic process. (Liu et al., 2010).Caspase-3 is one of the key

executioners of apoptosis, as it can be activated in both intrinsic and extrinsic pathway.

Activated caspase-3 has the potential of cleaving or degrading many key proteins such as

fodrin, nuclear lamins and the nuclear enzyme poly (ADP ribose) polymerase (PARP)

(Kaul et al., 2003). In our study, we demonstrated that Caspase-3 activity was

significantly up-regulated in KBrO3 treated group and pretreatment with B.monnieri

significantly restored the Caspase-3 activity.

p53 a Tumor Suppressor gene is implicated in cell cycle regulation, apoptosis and DNA

repair (Bowen et al., 2006).Both Caspase-3 and p53 are apoptosis associated proteins and

any change in their expression is related to instability of genome. Corsby et al., (2001)



demonstrated a modulation in cellular redox state upon KBrO3 exposure to activate p53

expression, a similar pattern of p53 expression was observed in our study and B.monnieri

supplementation inhibited this over-expression of p53.

Multiple genetic mutations cause irregular cell proliferation which is a crucial aspect of

carcinogenesis.ODC is the first and rate limiting enzyme in the biosynthesis of

polyamines, which are important for DNA synthesis and hence for cell proliferation.

ODC and the resulting polyamines are necessary for cellular proliferation, induction of

ODC is implicated in tumor promotion and cell transformation, and cultured tumor cells

often have high levels of ODC.

Thus, ODC is considered as an important marker tumor promotion and favorable target

for chemoprevention (Pegg et al., 1995). KBrO3administration in the present study led to

a marked elevation in ODC activity and [3H] thymidine incorporation in the kidney,

which was significantly ameliorated by pretreatment with B.monnieri. Thus, confirming

its role as potent antitumor promoting agent.

Many studies have revealed oxidative stress to be the major cause of the inflammation

stimulating the release of several proinflammatory cytokines. It also plays a significant

role in the development of renal nephropathy (Rivero et al., 2009). Redox status has also

been shown to influence NF-κB regulation and hence several genes involved in cell

transformation, proliferation, and angiogenesis. Although, relationship between ROS and

NF-κB is complex, ROS are believed to be implicated as second messengers in the

activation of NF-κB via TNF-α and other proinflammatory cytokines (Reuter et al.,

2010). Inhibition of NF-κB is a good strategy to control carcinogenesis and tumor

promotion. Since NF-κB is sensitive to ROS it is activated by strong oxidants like

KBrO3.Thus KBrO3 toxicity too can, at least to a certain extent, be controlled by NF-κB

inhibitors.

Further there is enough evidence that COX-2 inhibition represents a suitable target for

prevention of various proliferative diseases including cancer (Hull 2005). COX-2 is also

under the transcriptional regulation of NFκB by virtue of which it makes the environment

conducive for tumor promotion (Kaur et al., 2007). We have for the first time showed an



increase in expression of COX-2 in renal tissues following KBrO3 administration.

B.monnieri pretreatment significantly suppressed KBrO3 induced COX-2 expression.

PGE2, an important proinflammatory mediator whose synthesis is catalyzed by COX-2,

was also significantly elevated following KBrO3 exposure (Table 2). PGE2 is implicated

in inducing systemic inflammation and up-regulation the expression of important

cytokines such as IL-6 and in inducing COX-2 itself (Faour, et al., 2008). Thus PGE2

induction further aggregates the existing inflammation. PGE2 amelioration by B.monnieri

extract appears to be an important mechanism of its protective efficacy against renal

carcinogenesis.

NO (nitric oxide) is another important mediator in the pathogenesis of inflammatory

diseases (Federico et al., 2007). Thus, we also evaluated the effect of B.monnieri on renal

NO in KBrO3 induced renal toxicity (Table 2); KBrO3 exposure elevated the level of NO.

However, pretreatment with B.monnieri prevented KBrO3 induced NO production in a

concentration-dependent manner. Further, MPO is an enzyme formed mainly by

polymorphonuclear leucocytes, and is linked to the degree of neutrophil infiltration in a

given tissue (Sehirli et al., 2010). Following KBrO3 treatment, MPO activity was

markedly elevated (Table 2).This increase in MPO activity was significantly prevented

by B.monnieri administration, which is consistent to the histopathopathological findings.

In the present study, the biochemical alterations and the noticeable increase in BUN,

creatinine and LDH level, indicate proximal tubular dysfunction. The presence of tubular

damage was further confirmed by increased level of brush border marker; gamma

glutamyl transferase suggesting a direct toxic injury (Table 2). Renal morphologic

examination, which revealed the presence of tubular necrosis and proteinous casts further

confirmed renal tubular damage. These changes in histostructure of kidneys were

successfully restored by higher dose of B.monnieri (Figure 3).

The results of our study infer that pretreatment with B.monnieri inhibits KBrO3 induced

renal carcinogenesis. Oral supplementation of B.monnieri prior to KBrO3 exposure

resulted in a noticeable decline in protein expression of COX-2 and p53, secretion of

proinflammatory cytokines, ODC activity, [3H]-thymidine incorporation into DNA, all of



which are established markers of inflammation and tumor promotion. In addition, we

have also demonstrated that B.monnieri helps to maintain antioxidant armory, alleviates

serum toxicity markers and also suppresses activation of redox active transcription factor:

NFκB. These results suggest B.monnieri extract to be a capable candidate for

chemoprevention of renal carcinogenesis since it restrains several biomarkers of

inflammation, Oxidative stress, Apoptosis and tumor promotion in an animal model.



Figure 1 In-Vitro Assessment of B. Monnieri

Assessment of antioxidant potential of Q. Infectoria in terms of Lipid Peroxidation, DNA

damage reducing potential assay and DPPH assay.



Figure 2 Immunohistochemistry of COX-2, Representative photomicrographs of

COX-2 determined by immunohistochemistry

(B) There is Over expression of COX-2 as represented by black arrows was observed in

KBrO3 treated group. (A) While control group showed almost negligible staining of

COX-2 positive cells (C) There was partial inhibition of COX-2 expression as shown by

weak immunostaining of rat kidneys section treated with lower dose of B.monnieri (100

mg/kg b.wt.). (D) In group treated with higher dose of B.monnieri, (200 mg/kg b.wt.)

there was very slight staining of COX-2. (40X magnification)



Figure 3 Representation of immunehistochemical detection of p53 in renal sections

Over expression of p53 as represented by dark brown DAB positive was observed in

KBrO3 treated group (B), while in control group there was no staining of p53 positive

cells (A). B.monnieri treatment in lower dose (100 mg/kg b.wt.) showed lesser number of

p53 positive cells (C) and group treated with higher dose (200 mg/kg b.wt.) of

B.monnieri presented very moderate staining of p53 (D). (40X magnification)



Figure 4 Effect of B.monnieri treatment on renal histological alterations caused by KBrO3

application

(A) represents control group, slide (B) represent KBrO3 treated group. (C) and (D) are

B.monnieri + KBrO3 groups (100 and 200mg/kgb.wt. respectively), (D) There were no

changes observed in renal architecture. Normal renal morphology can be seen clearly. (B)

Massive tubular necrosis as represented by arrows and proteinous casts is observed,

infiltration of the interstitial cells along with acute nephritis can also be observed. Overall

renal morphology is damaged by KBrO3 administration. (C-D) Slides show that

B.monnieri maintains the renal architecture and minimizes the infiltration of

inflammatory cells and nephritis. (40X magnification)



Figure 5 Inhibitory effect of B. Monnieri on KBrO3 induced expression of NFĸB in rat

renal tissues:

Following exposure to KBrO3, protein expressions were determined by

Immunohistochemical detection of NFĸB in renal section. (A) Control animals with no

expression of NFĸB, (B) Brown stain represent NFĸB overexpression in KBrO3-treated

rats (C) 100mg/kg BW B. Monnieri oral supplementation decreases the expression of

protein (D) 200 mg/kg BW B. Monnieri oral supplementation shows complete inhibition

of NFĸB expression. (40X magnification)



Figure 6 Effect of prophylactic treatment of B.monnieri on caspase-3 activity in the Kidney

of KBrO3 administered rats:

Values are expressed as mean ± SE of six animals per group. Caspase-3 activity was

significantly increased in the KBrO3 Group (***p < 0.001) as compared to Control

Group. Lower dose (100 mg/kg b.wt.) of B.monnieri (BM1+KBrO3) exhibited significant

(##p < 0.01) change in caspase-3 activity as compared to KBrO3 treated group.

Pretreatment with higher dose (200 mg/kg b.wt.) of B.monnieri (BM1+KBrO3)

significantly (###p < 0.001) decreased the caspase-3 activity as compared to KBrO3

alone treated group.



Figure 7 Effect of B.monnieri pretreatment on kidney ODC in KBrO3 induced

nephrotoxicity:

ODC level was significantly (***p< 0.001) elevated in KBrO3 treated group II as

compared to group I control. Pretreatment with B.monnieri attenuates the ODC level

significantly in the KBrO3 + D1 & KBrO3 + D2 groups in comparison with KBrO3 (alone)

treated group II (##p < 0.01, ###p < 0.001).



Figure 8 Effect of B.monnieri pretreatment on kidney renal DNA synthesis in

KBrO3 induced nephrotoxicity:

KBrO3 administration strongly increased thymidine incorporation (***P<0.001) as

compared to control. Pretreatment with B.monnieri significantly restores this level of

thymidine to normal at higher dose (KBrO3 + D2) as compared to KBrO3 (alone) treated

group (#p < 0.05).



Table.1 Results of pretreatment of B.monnieri on antioxidant enzymes like GSH,

GST, GR and GPX on KBrO3 induced renal redox imbalance.

Results represent mean ± SE of six animals per group. Results obtained are significantly

different from Control group (***P < 0.001). Results obtained are significantly different

from KBrO3 treated group (#P < 0.05), (##P < 0.01), (ns P= not significant) and

(###P<0001).BM= B.monnieri; D1= 100mg/kg/b wt; D2 = 200mg/kg/b wt.

Treatment

regimen

per group

GSH

(n mol GSH

/g tissue)

GST conjugate

formed/min/mg

protein)

GR

(n mol NADPH

Oxidized/min/

mg protein)

GPX

(n mol

NADPH

Oxidized/min/

mg protein)

Group I (control) 0.46±0.03 137.4±8.8 201.9±7.71 285.3±13.50

Group II

(KBrO3 only) 0.24±0.01*** 74.61±2.62** 110.9±2.59*** 158.8±9.92***

Group III

(KBrO3+B M D1) 0.38±0.01### 96.30±5.64ns 150.0±6.81# 214.0±5.55#

Group IV

(KBrO3+ + BM

D2)

0.39±0.01### 127.9±7.84# 165.8±15.98## 219.6±16.83##

Group V

(only BM D2) 0.48±0.02 142.1±18.19 195.7±6.54 286.4±5.74



Table.2 Results of pretreatment of B.monnieri on parameters like Catalase, MDA,

XO and SOD on KBrO3 induced renal biochemical imbalance.




(###P<0001). BM =B.monnieri; D1= 100mg/kg/b wt; D2 = 200mg/kg/b wt.

Treatment

regimen

per group

Catalase

(nmol H2O2

consumed/min/mg

protein)

MDA

(nmol MDA

formed/g tissue)

XO

(µg of uric acid

formed/min mg

protein)

SOD

(n moles p-

nitroaniline/min/mg

protein)

Group I (control) 0.46±0.03 2.93 ± 0.11 0.206±0.02 174.9 ± 4.93

Group II (KBrO3

only) 0.24±0.01*** 5.65 ± 0.16*** 0.431 ± 0.03*** 116.6± 6.64***

Group III (

KBrO3+B M D1) 0.38±0.01### 5.09 ± 0.35ns 0.370 ± 0.05ns 134.4 ± 2.94#

Group IV(

KBrO3+ + BM

D2)

0.39±0.01### 3.48 ± 0.12### 0.229 ± 0.04# 151.6 ± 2.83###

Group V (only

BM D2) 0.48±0.02 2.83 ± 0.16 0.212 ± 0.02 176.9 ± 4.71



Table.3 Results of pretreatment of B.monnieri on Toxicity Markers like BUN,

Creatinine, LDH and GGT on KBrO3 induced enhancement


different from Control grou (***P < 0.001). Results obtained are significantly different

from KBrO3 treated group (#P < 0.05), (##P < 0.01) and

(###P<0.001).BM= B.monnieri; D1= 100mg/kg/b wt; D2 = 200mg/kg/b wt.

Treatment regimen

per group

BUN

(mg / 100 ml)

IU/L

Creatinine

(mg / 100

ml/IU/L

LDH

(n mol NADH

oxidized / min/ mg

protein)

γ-GGT

(nmoles p

nitroaniline

min/mg protein)

Group I (control) 19.51±1.84 1.89±0.06 222.2±14.19 595.3 ± 8.347

Group II (KBrO3

only) 41.49±2.85* 3.62±0.37* 545.90±30.77*** 817.8± 18.51***

Group III (KBrO3+

BM D1) 28.97±1.01## 2.30±0.23## 374.5±26.84## 732.7 ± 23.62#

Group IV

(KBrO3+BM D1) 26.32±0.82## 1.94±0.13## 346.02±38.02### 610.1 ± 6.61###

Group (only BM D2) 19.14±2.20 1.92±0.04 203.1±10.53 593.7 ± 21.69



Table.4 Results of modulatory effect of B.monnieri on inflammatory and

proinflammatory parameters like TNF-α, PG E2, Nitric oxide and Neutrophil infiltration

on KBrO3 induced renal redox imbalance.




(###P<0001).BM= B.monnieri; D1= 100mg/kg/b wt; D2 = 200mg/kg/b wt.

Treatment regimen

per group

TNF-α

(TNF-α

pg/ml)

PG E2

(PGE2 pg/ml)

NO

(m moles of

nitrite/mg of

tissue)

MPO

(units of MPO

activity/min/mg

protein)

Group I (control) 311.9±39.2 49.20±4.44 19.60±1.12 0.114±0.01

Group II (KBrO3

only) 718.3±24.1*** 155.6±10.5*** 49.40 ± 2.50*** 0.248 ± 0.02***

Group III ( KBrO3+B

M D1) 545.2±58.2# 102.6±9.66## 34.60± 2.18### 0.198± 0.02ns

Group IV( KBrO3+ +

BM D2) 460.7±52.9## 86.80±15.8### 26.40 ± 2.75### 0.174± 0.01##

Group V (only BM

D2) 307.1±41.3 46.70±4.43 22.80 ± 0.80 0.106 ± 0.01

chapter 7 bacopa - information and library network...

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