Inter. J. of Phytotherapy / Vol 5 / Issue 2 / 2015 / 70-75.

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e - ISSN - 2249-7722

Print ISSN - 2249-7730

International Journal of Phytotherapy

www.phytotherapyjournal.com

ANTIHYPERGLYCEMIC AND ANTIHYPERLIPIDEMIC

ACTIVITIES OF ETHANOLIC EXTRACTS OF CASSIA SOPHERA

(L.) IN ALLOXAN INDUCED DIABETES MICE

Chand Sultana Khatun1, Bytul Mokardesh Rahman

3, Naznin Ara Khatun

3, A.K. Azad

1,

Ashraf Ali2 and Mir Imam Ibne Wahed*

3

1Department of Pharmacy, Bangladesh University, Dhaka-1207, Bangladesh. 2Department of Pharmacy, Southeast University, Dhaka-1212, Bangladesh.

3Department of Pharmacy, Rajshahi University, Rajshahi-6205, Bangladesh.

INTRODUCTION

Diabetes is a global disease with huge adverse

impacts on health and mortality particularly of

cardiovascular disorders. Diabetes mellitus (DM) is the

major clinical disorder affecting nearly 10% of the

populations all over the world [1]. The prevalence of the

DM is increasing rapidly in the developing countries than

in the developed country. There are an estimated 246

million people with diabetes in the world, of whom about

80% reside in developing countries [2]. Patient with

diabetes have an increased risk of coronary heart disease,

peripheral vascular disease, strokes and may account for

more than 65% death among people with diabetes [3-5].

Multiple pathophysiologic mechanisms play a role in the

risk of cardiovascular events in the metabolic syndrome

including glucose intolerance, hyperglycemia,

hypertension, dyslipidemia, atherosclerosis that are

caused primarily by insulin resistance [6-7]. In particular

hyperglycemia may contribute to diabetic complications

Corresponding Author:- Mir Imam Ibne Wahed Email: wahed_mir@ru.ac.bd

ABSTRACT

The aim of the investigation was to investigate the hypoglycemic and hypolipidemic activity of ethanolic

extracts of different parts (fruits and stems) from the plant Cassia sophera in alloxan-,induced diabetic mice. 20 mice were used for a week-long anti-hyperglycemic test. Lipid profile analysis was performed by estimating total

cholesterol (TC), triglyceride (TG), phospholipid and liver glycogen concentration on 20 mice after week-long

antihyperglycemic tested mice. Metformin Hcl (150mg/kg body weight, op.) was used as a standard antidiabetic

agent. The result indicated that after oral ingestion of Cassia sophera(CS) extracts for seven days in diabetic mice,

Group-DCF and Group-DCS mice showed significantly (p

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by altering vascular cellular metabolism, vascular matrix

molecules and circulating lipoproteins. However, an

abnormal lipid profile was found to a more significant

risk factor than either hypertension or diabetes alone [8].

The burden of death and disability from diabetes and its

related complications remain great due to their therapeutic

failure as well as the potential for induction of

hypoglycemia [9]. Traditional medicines are used to

reduce blood glucose level as well as have beneficial

effects on complication of diabetes [10]. Therefore,

natural agents having hypoglycemic and hypolipidemic

properties would be better for the management of

diabetes.

Cassia sophera Linn. (CS) belonging to the

family Caesalpiniaceae, an annual or perennial shrub or

under shrub with 18-23cm grows in waste lands all over

the country. It is known as Sennasophera in English and

Kasondi in Bengali.Cassia sophera leaves contain a

flavanol c-glycoside and sennosides and root barks

contain anthraquinones, chrysophanol and physicion and

betasitosterol and flowers contain anthraquinone [11]. A

novel cycloartanetriperpene glycoside obtained from the

seeds of Cassia sophera [12]. Pharamacological

investigation revealed that Cassia sophera is effective in

the treatment of asthma, acute bronchitis, cough, diabetes

and convulsions of children [13-16].Cassia sophera plant

extracts inhibiting molecular interactions between nuclear

factors and target DNA sequences mimicking NF-kappa

B binding sites. It is insecticidal and known to produce

vertebrate toxicity [17]. However, there are no reports on

the anti-diabetic activity of Cassia sophera have been

found.

MATERIALS AND METHODS

Plant materials

Fresh plants (stems & fruits) of Cassia sophera

Linn.were collected from Rajshahi in August 2013 and

the plant authenticity was confirmed from the Bangladesh

National Herbarium, Mirpur, Dhaka. A voucher specimen

no. 33166 is maintained in our laboratory for future

reference.

Extraction

The plant (stem & fruits) collected were washed

and sun dried under shadow for several days. The

powdered plant leaves were extracted with 96% ethanol at

room temperature. The bottle were kept at room

temperature and allowed to stand for several 7-10 days

with occasional shaking and stirring. The extracts thus

obtained were filtered through cotton and then through

filter paper (Whatman Filter Paper No. 1). The filtrate

was defatted with petroleum ether for several times. Then,

the defatted liquor was allowed to evaporate using rotary

evaporator at temperature 40-45C. Thus the highly

concentrated crude extracts were obtained.

Drugs and chemicals

The active drug, Metformin hydrochloride was

the generous gift samples from Square Pharmaceuticals

Ltd; Pabna Bangladesh. Total cholesterol (TC) and

triglyceride (TG) wet reagent diagnostic kits were in the

products of Crescent diagnostic kits. Alloxan was

purchased from Sisco Research Laboratories Pvt. Ltd.

Mumbai, India. All other chemicals and solvents were of

analytical grade.

Animals

Nine weeks old male Swiss Albino mice (weight,

25-28g) were purchased from ICDDRB, Dhaka,

Bangladesh and housed in animals cages under standard

environmental conditions (22-25C, humidity 60-70%)

with water ad. libitum. The animals used in this study

were cared in according to guidelines of animal

experiment.

Phytochemical screening

Phytochemical analysis was performed

according to the standard methods described by Nayek

and Pereira [18].

Anti hyperglycemic test

Anti hyperglycemic test was performed

according to standard method. All mice were divided into

four groups and each group comprised of five mice.

Groups were Group-DC (Diabetic Control groups

receiving vehicle 0.5% methyl cellulose), Group-DS

(Diabetic Standard group mice received Metformin HCl,

150 mg/kg), Group-DCF and Group-DCS (Diabetic

Cassia Fruit and Diabetic Cassia Stem group mice

received ethanolic extracts of Cassia fruits and stems 300

mg/Kg body weight, dissolved in 0.5% methyl cellulose,

respectively). After fasting 16 hours, diabetes was

induced to all groups by intra-peritoneal injection (IP) of

alloxan monohydrate (100 mg/kg) dissolved in saline.

After 72 hours, blood glucose levels were measured from

tail-vein blood of all groups by Glucometer considered as

0 day and blood glucose level higher than 11.5 mmol/l

considered as diabetic. 0.5% methyl cellulose, standard

drug metformin and extracts (300 mg/kg) were

administered once daily for seven days to respective mice

groups. Blood glucose content was measured after 1st, 3

rd

and 7th

days by Glucometer.

Blood and organs sampling

The body weight of mice of each groups were

measured before and after week-long antihyperglycemic

tested with drugs. The mice were sacrificed by

anesthetizing with pentobarbital (5mg/kg, i.p.), blood

samples were withdrawn from aorta of heart using a

syringe and kept into an EDTA containing tube. Heart,

liver and kidney were excised and cleaned of the

surrounding tissues. The organ weights were measured

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immediately and the organ weight to body weight ratio

were calculated. Finally, the blood and tissue samples

were preserved in refrigerator at -40C for biochemical estimations.

Estimation of serum cholesterol, triglyceride and

phospholipids in diabetic mice

Serum samples were obtained by centrifugation

of blood at 4000 rpm for 10 minutes. The concentration of

TC and TG were measured by UV-spectrophotometer,

using wet reagent diagnostic kits according to the

manufacturers protocol. Total lipid was extracted from the liver and kidney tissue according to the method of

Folch et al [19]. Phospholipids were estimated by the

method of Bartlette by digestion with perchloric acid and

the phosphorous liberated was estimated by the method of

Fiske and Subbarow [20-21].

Estimation of glycogen concentration in liver of

diabetic mice

The liver glycogen content was determined

according to the method described by Tarnoky K. et. al.

Briefly, it utilizes the o-toluidine-glucose coupling

reaction for the estimation of glycogen after

trichloroacetic acid (TCA) extraction, precipitation by

alcohol and hydrolysis [22]

STATISTICAL ANALYSIS

The results were expressed as mean Standard

Error of Mean (SEM). Statistical analysis was performed

by using ANOVA followed by Tukeys test using Graph pad Prism Software version 5.03. P values

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Table 3. Effect of CS extracts on body weight and body weight to organ weight ratio in diabetic mice

Group

Initial

Body weight (g)

Final

Body weight (g)

HW/BW

(g/kg)

LW/BW

(g/kg) KW/BW (g/kg)

Group-DC 281.40 35.51.20 0.770.12 8.51.5 1.70.21

Group-DS 321.7 272.12 0.310.5 4.110.11** 0.720.16

Group-DCF 27.62.12 28.02.12 0.420.19 5.40.10* 0.870.54

Group-DCS 30.562.8 290.70 0.440.07 5.20.10* 0.300.04

Values were expressed in Mean SEM. Control group received 0.5% Methyl cellulose and standard group received

Metformin 150 mg/kg. *p

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plants earlier [25]. The increase concentration of free fatty

acid in liver and kidney may be due to lipid breakdown

and this may cause increased generation of NADPH

dependent microsomal lipid proxidation during diabetes.

As a result liver and kidney phospholipids were increased

in diabetic mice [26]. Administration of CS extracts

significant decrease the level of tissue free fatty acids and

phospholipids. Induction of diabetes with alloxan caused

decrease in hepatic glycogen, which could be attributed to

the decrease in the availability of the active form of

enzyme glycogen synthetase probably because of low

levels of insulin [27]. In this study, CS extracts restored

the depressed hepatic glycogen levels possibly by

increasing insulin (figure 1). Decreased activities of the

enzymes involved in glucose homeostasis in liver and

kidney such as hexokinase has been reported in diabetic

animal resulting in depletion of liver and muscle glycogen

content.

CONCLUSION

Based on results of present study, we conclude

that the plant extracts of Cassia sopherafruits and

stemspossess blood glucose and lipid lowering activities.

However, further studies are needed to isolate active

compounds responsible for these pharmacological

activities and also necessary to examine underlying

mechanism of antidiabetic and lipid lowering effects of

Cassia sophera.

ACKNOWLEDGEMENT The authors would like to express thanks, to the

Department of Pharmacy, Rajshahi University, Rajshahi,

Bangladesh for providing lab facilities and International

Centre for Diarrhoeal Disease Research, Bangladesh

(ICDDR, B) for supplying animals (mice) for the research

work.

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