a comparative study of in-vitro thrombolytic activity and anti-inflammatory activity of methanolic...

Presented byExamination Roll No. # 12810

Registration No: 776Session : January 2012

Phytochemical and Pharmacological Research Laboratory

Department of PharmacyUniversity of Science & Technology Chittagong (USTC)

Foy’s Lake, Khulshi, Chittagong-4202, BangladeshAugust, 2016

A comparative study of In-vitro Thrombolytic and Anti-inflammatory activity of methanolic leaves extract of Blumea lacera (Asteraceae)

& Acanthus ilicifolius (Acanthaceae).

The plants that possess therapeutic properties or exert beneficial pharmacological effects on the

animal body are generally designed as ‘medicinal plants’. Although there are no apparent

morphological characteristics in the medicinal plants that make them distinct from other plants

growing with them , yet they possess some special qualities or virtues that make them medicinally

important.

About 80% of the population in developing countries relies on traditional plant based medicines for their

primary health care needs.

INTRODUCTION

Guggul Tree Gugulsterone Pacific Yew Paclitaxel (Taxol)

The Aim of work includes the following:

Investigation of in-vitro thrombolytic of Blumea lacera. Investigation of in-vitro thrombolytic of Acanthus

ilicifolius. Investigation of anti-inflammatory activity of Blumea

lacera. Investigation of anti-inflammatory of Acanthus ilicifolius. Compare the result of in-vitro thrombolytic activity of

Blumea lacera and Acanthus ilicifolius. Compare the result of anti-inflammatory activity of

Blumea lacera and Acanthus ilicifolius.

AIM OF WORK

Blumea lacera .Taxonomy of Blumea lacera :

Kingdom: Plantae Subkingdom: Tracheobionta Division: Magnoliophyta Class: Magnoliopsida Subclass: Asteriadae Order: Asterales Family: Asteraceae Genus: Blumea Species: lacera

PLANT PROFILELocal Name: Bara Kukshima,

Kukursunga, Shealmotra, Shealmoti.

Using Information: Plant is astringent, stomachic, antispasmodic, antipyretic and diuretic; cures bronchitis, fevers and burning sensation. Leaf juice is anthelmintic,

astringent, febrifuge, stimulant and diuretic; mixed with pepper, it is given in bleeding piles. Roots mixed with pepper are given in cholera and along with rhizomes of

Cyperus rotundus given in dysentery.

Chemical structure of some active phyto-constituents of Blumea lacera

CineoleFenchone

Campesterol

α-amyrin

ß-sitosterol

Hentriacontane Hentriacontanol

Lupeol Prenylated phenolic glycosides

Acanthus ilicifolius.Taxonomy of Acanthus ilicifolius:

Kingdom: Plantae Phylum: Tracheophyta

Division: Magnoliophyta Class: Magnoliopsida

Subclass: Asteriadae Order: Scrophulariales Family: Acanthaceae Genus: Acanthus Species: ilicifolius

PLANT PROFILELocal Name: Hargoza, Harkuch

Kanta, Kotki.

Using Information: Root possesses expectorant properties; used in cough and asthma. Leaves are used as fomentation in rheumatism and neuralgia. As a

stimulant the decoction of plant with sugarcandy and cumin is given in dyspepsia.

Chemical structure of some active phyto-constituents of Acanthus ilicifolius

Acanthicifolin Quercetin ß-sitosterol

Oleanolic Acid Lupeol Apeganine-7-glucoronide

The methanol extract of the whole plant of Blumea lacera (Burn.f.) DC. (BLME)

has been subjected to preliminary screenings for phytoconstituents and antipyretic,

analgesic and anti-inflammatory activities was assessed By (Md. Abul Khair et al)

Hence, Acanthus ilicifolis is considered to be rich sources of steroids, triterpenoids,

saponins, flavonoids, alkaloids, and tannins. Present review article is an attempt to

cover recent developments in phytochemical and pharmacological potential of drug.

Traditionally, the plant has been used for dyspepsia, paralysis, asthsma, headache,

rheumatism, and skin diseases (Dharya Singh et al)

To investigate the antimicrobial activity of Acanthus ilicifolius against the skin

infecting bacterial and fungal pathogens (Chinnavenkataraman Govindasamy et al)

Literature Review

The essential oil of Blumea lacera DC. leaves was analysed by GLC and GC-MS.

Thymoquinol dimethyl ether was found to be the main constituent. Other major

compounds included β-caryophyllene, α-humulene and E-β-farnesene.

Thymoquinol dimethyl ether and precocene (I. Laakso et al)

To investigate the antimicrobial activity of Acanthus ilicifolius against the skin

infecting bacterial and fungal pathogens (Chinnavenkataraman Govindasamy et al

Campesterol was isolated from Blumea lacera DC. The structure was elucidated on

the basis of the data of NMR, MS, IR. ( Chakravarti et al)

Continue…..

Research Protocol Collection, drying

& grinding the leaves of Pothos scandens L.

Collection, drying & grinding the leaves of

Blumea lacera & Acanthus ilicifolius

Major Methanolic Extract

A comparative study of In-vitro Thrombolytic activity and Anti-inflammatory activity of methanolic leaves extract ofBlumea lacera and Acanthus ilicifolius

Selection of plant material:

Blumea laceraFamily : Asteraceae

Acanthus ilicifoliusFamily : Acanthaceae

Collection of plant material:

The fresh Leaves of Blumea lacera and Acanthus ilicifolius were collected from Chittagong Hill Tracts at 25th May,2016 by using hand labor with appropriate manner.

Identification of plant material:

The leaves of Blumea lacera and Acanthus ilicifolius were identified by using standard taxonomical methods, supervised by Dr. Shaikh Bokhtear Uddin, Associate Professor, Department of Botany, University of Chittagong, Bangladesh and was taken voucher specimen from a Chittagong University Herbarium.

Materials & Method

Extraction of Leaves

Collection & drying

Leaves powder of Blumea lacera and Acanthus ilicifolius

Grinding

Methanol addition

Methanolic extract

Collection of crude extract

Allowed to stand for solvent evaporation

TEST NAME Inferences for Blumea lacera

Inferences for Acanthus ilicifolius

Test for Alkaloids + +

Test for glycoside + +

Test for Tannins + +

Test for Flavonoids + +

Test for Steroids + +

Test for Saponins (Shake test)

- +

Screening Test

Test for Steroid Test for Glycoside

Test for Saponins Test for Tanins

Test for Flavanoids

Test for Alkaloids

Acanthus ilicifoliusBlumea lacera

Test for TanninsTest for Saponins

Test for GlycosidesTest for Steroids

Test for Flavanoids Test for Alkaloids

Thrombolytic activity

% of clot lyses = lysis beforeclot ofWeight lysisafter clot ofWeight

100

Results of thrombolytic activity of Blumea lacera and Acanthus ilicifolius

Sample % of Clot lysis

Distilled Water (Negative Control) 4.6

Blumea lacera ( Methanol extract) 49.27

Acanthus ilicifolius ( Methanol extract) 38.25

Streptokinase (Positive Control) 69.34

Figure : % clot lysis by Water, methanolic extract and streptokinase

Discussion :The mean % of clot lysis for water and streptokinase was found 4.6% and 69.34% respectively. The mean percent clot lytic activity of methanolic leaves extract of Blumea lacera was found 49.27% .On the other hand, The mean percent clot lytic activity of methanolic extract of Acanthus ilicifolius was found 38.25%.It shows Blumea lacera and Acanthus ilicifolius leaves both have moderate thrombolytic activity.

0

10

20

30

40

50

60

70

80

4.6%

38.25%

49.27%

69.34%

Effect of methanolic extract of Blumea lacera & Acanthus ilicifolius

DW MEBL MEAI Streptokinase

% o

f Clo

t Lys

is

Anti-inflammatory activity

control of absorbancesample of absorbance% inhibition = 1- x 100

Absorbance(at 660nm)

Mean Absorbance

% MIPD % MPD

Control(Methanol)

1. 0.095 0.095 0

1002. 0.097

3. 0.094Standard(Acetyl salicylic acid 0.1gm)

1. 0.031 0.027 71.58

28.422. 0.0343. 0.017

MEBL(1 mg/ml)

1. 0.037 0.0336 64.56 35.452. 0.031

3. 0.033MEAI(1mg/ml)

1. 0.023 0.0246 45.27 21.192. 0.027

3. 0.024

Results of Anti-inflammatory activity of Blumea lacera and Acanthus ilicifolius

From the above study it was observed that leaves of Blumea lacera and Acanthus ilicifolius as an herbal medicine in the treatment of inflammatory diseases which % inhibition of protein denaturation is quite significant with standard acetyl salicylic acid.

DISCUSSION :

0

20

40

60

80

100

120

0

71.5864.56

45.27

100

28.4235.45

21.19

%MIPD

%MPD

Figure: Mean inhibition of protein denaturation vs methanolic fraction of methanolic leaves extract of Blumea lacera and Acanthus ilicifolius

The results from the experiment confirmed that these leaves extracts shows significant thrombolytic and anti-inflammatory activities.The comparison of In-vitro Thrombolytic activity between Blumea lacera and Acanthus ilicifolius methanolic leaves extract are, “ Blumea lacera have 49.27% clot lysis and Acanthus ilicifolius have 38.25% clot lysis with 4.6% water (negative control) and 69.34% streptokinase (positive control) ”.

On the other side, the comparison of Anti-inflammatory activity between Blumea lacera and Acanthus ilicifolius methanolic leaves extract are, “ Inhibition of protein denaturation of methanolic extracts of Blumea lacera is (64.56±0.00176)% and the inhibiton of protein denaturation of methanolic extracts of Acanthus ilicifolius is (45.27±0.00143)% is significant with Acetyl saliylic acid(Standard).

However, further studies would be necessary to evaluate the contribution of other substances for the activities showed and elucidate the exact mechanism behind these observed activities as it still remains to be determined which components were exactly responsible for these effect.

Conclusion

Abdul Ghani, Practical phytochemistry: including methods of phytochemical studies, ISBN: 984-32-2310-1.

DeSmet P.As. (1997), The role of plant derived drugs and herbal medicines in healthcare, Drugs,Vol. 54,pp. 108.

Ghani A. Medicinal plants for drug development, potentiality of medicinal plants of Bangladesh. Tenth Asian symposium on medicinal plants, 2000; 18-23: 39.

Shaker A. Mousa. Anticoagulants, Antiplatelets, and Thrombolytics. Methods in Molecular medicine TM. vol.93.

Medicinal plants of Bangladesh by Dr. Shaikh Bokhtear Uddin. www.mpbd.com.

Caius. J.F. 1986. The Medicinal and Poisonous Plants of India. Scientific Publ. Jodhpur, India. p. 323-325.

Handin RI 92005). “ Chapter 53: Inflammation”. In Kasper DL, Braunwald E, Fauci AS, et al. Harrison’s Principles of Internal Medicine (16th edition).

REFERENCES

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