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

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International Journal of Phytotherapy

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ANTIHYPERGLYCEMIC AND ANTIHYPERLIPIDEMIC

ACTIVITIES OF ETHANOLIC EXTRACTS OF CASSIA SOPHERA
(L.) IN ALLOXAN INDUCED DIABETES MICE
Chand Sultana Khatun1, Bytul Mokardesh Rahman3, Naznin Ara Khatun3, A.K. Azad 1,
Ashraf Ali2 and Mir Imam Ibne Wahed*3
1

Department of Pharmacy, Bangladesh University, Dhaka-1207, Bangladesh.

2
Department of Pharmacy, Southeast University, Dhaka-1212, Bangladesh.
3
Department of Pharmacy, Rajshahi University, Rajshahi-6205, Bangladesh.

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<0.05) reduced bloodglucose levels 47.23% and62.13%
respectively at the dose 300 mg/kg body weight when compared to Group-DC mice. Short-term treatment with
extracts had no effect on body weight to organ weight ratio; however, it significantly lowered liver weight in GroupDCF and Group-DCS. Administration of extracts greatly reduced both the serum cholesterol and triglycerides and
phospholipids levels in Group-DCF and Group-DCS. Liver glycogen content significantly restored in Group-DCF
and Group-DCS compared to Group-DC mice. The present investigation established the pharmacological evidence
to support the folklore claim and that the plant parts have anti-diabetic and hypo-lipidemic potential.
Key words: Cassia sophera(CS), Total Cholesterol (TC), Triglyceride (TC), Phospholipids and Liver Glycogen.

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

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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 1 st, 3rd
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 <0.05, p<0.01
and p<0.001 were considered as statistically significant.
RESULTS
Antihyperglycemic effect of CSextracts in diabetic
mice
Hypoglycemic test was performed with extract
showed more improvement compared with Group-DC
table-2). After 7 days mice treated with extracts in Group-

DCF and Group-DCS (300 mg/kg body weight), glucose

level were significantly lowered 47.23% and 62.13%
respectively showed in table 2.
Effect of CS extracts on body weight, organ weight
(heart, liver and kidney) and organ weight to body
weight ratio changes
Table 3 shows no significantly changes in the
body weight among experimental animal after seven days
treatment. The results revealed that heart weight to body
weight and kidney weight to body weight ratio did not
altered significantly in experimental mice compared to
Group-DC. Although the liver weight to body weight
ratio were severely altered or increase in Group-DC, after
treatment. It was significantly decreased in Group-DCF
and Group-DCS compared to the Group-DC mice
although total food intake was not different.
Effects of CS extracts on total cholesterol, triglyceride
and phospholipid in diabetic mice.
Comparison of serum lipid contents in control
group and experimental groups of mice are shown in table
4. Administration of CS extracts and metformin to
diabetic mice a significant decrease (p<0.05) in the levels
of total cholesterol, triglycerides, and phospholipids
observed in Group-DS, Group-DCF and Group-DCS
compared to Group-DC. Administration of CS extracts
and metformin mice tend to bring the levels of hepatic
lipids to near normal level.
Effects of CS extracts on liver glycogen level in
diabetic mice
In this study the level of glycogen in liver was
increased in Group-DS, Group-DCF and Group-DCS
compared to Group-DC. Treatment of diabetic mice with
metformin and experimental groups significantly (p<0.05)
improved the level of glycogen content compared to
Group- DC as shown in figure 2. Although Group-DS
gave more significant result than Group-DCF and GroupDCS.

Table 1.Phytochemical screening test result of CS extracts

Plant Extrats
Alkaloids
Saponins
Flavonoids
Cassia fruits extract
+
+
+
Cassia stems extract
+
+
+
+ = presence, - =Absence

Tanins
+
+

Triterpenoids
+
+

Table 2.Anti hyperglycemic effect of CS extracts in diabetic mice

Groups
0 day
1st day
3rd day
7th day
19.51.4
18.52.1
18.82.1
18.752.5
Group-DC
17.751.35
13.31.4
10.31.0*
5.71.1***
Group-DS
15.350.69
14.22.2
13.32.1
8.11.5*
Group-DCF
18.752.5
15.71.1
11.11.2*
7.11.6**
Group-DCS
Values were expressed in Mean SEM. Control group received 0.5% Methyl cellulose and standard group received
Metformin 150 mg/kg. *p<0.05, **p<0.01, and ***p<0.001 indicate significant changes compared with diabetic control.

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Table 3. Effect of CS extracts on body weight and body weight to organ weight ratio in diabetic mice
Initial
Final
HW/BW
LW/BW
KW/BW (g/kg)
Group
Body weight (g) Body weight (g)
(g/kg)
(g/kg)
281.40
35.51.20
0.770.12
8.51.5
1.70.21
Group-DC
321.7
272.12
0.310.5
4.110.11**
0.720.16
Group-DS
27.62.12
28.02.12
0.420.19
5.40.10*
0.870.54
Group-DCF
30.562.8
290.70
0.440.07
5.20.10*
0.300.04
Group-DCS
Values were expressed in Mean SEM. Control group received 0.5% Methyl cellulose and standard group received
Metformin 150 mg/kg. *p<0.05 and **p<0.01 indicate significant changes compared with diabetic control.
Table 4. Levels of serum total cholesterol, triglycerides and Phospholipids in diabetic mice after treatment with
extracts or drugs.
Groups
Total Cholesterolmg/dl
Triglyceridesmg/dl
Phospholipidmg/dl
1801.75
2042.4
173.5 13.4
Group-DC
86.61.45*
85.562.52*
91.7 8.5*
Group-DS
100.571.74*
101.762.11*
138.5 6.4*
Group-DCF
88.8 2.44*
90.72.04*
128.11.8*
Group-DCS
Values were expressed in Mean SEM. Control group received 0.5% Methyl cellulose and standard group received
Metformin 150 mg/kg. *p<0.05 indicate significant changes compared with diabetic control group.
Fig 2. Effects of CS extracts on the liver glycogen level compared to diabetic control

Values were expressed in Mean SEM. Control group received 0.5% Methyl cellulose and standard group received
Metformin 150 mg/kg. *p<0.05 and **p<0.01 indicate significant changes compared with diabetic control.
DISCUSSION
The
experiment
showed
that
the
antihyperglycemic experiment of extracts on diabetic
mice showed lower blood glucose level (table 2). In this
experiment, Group-DCF and Group-DCS has a significant
decrease in blood glucose concentration 47.23% and
62.13% respectively after 7th days treatment. Extracts
may have the properties to stimulate -cell for the
secretion of insulin and are most effective for controlling
diabetes due to presence of hypoglycemic alkaloid,
saponin and flavonoid (table-1). The extracts might be
promoting glucose uptake and metabolism or inhibiting
hepatic gluconeogenesis [23]. Short-term treatment with
extracts had no effect on body weight to organ weight
ratio; however, it significantly lowered liver weight in
Group-DCF and Group-DCS compared to Group-DC
(table 3).

In hyperglycemic mice there was a significant

increased in lipids (total cholesterol and triglycerides).
The most common lipid abnormalities in diabetes are
hypercholesterolemia
and
hypertriglyceridmia.Oral
administration of CS extracts resulted in a significant
reduction of serum lipids level in mice, viz total
cholesterol and triglycerides. Flavonoids are known for
their diverse activities including hypolipidemic and
antioxidant activity [24]. CS extracts contain flavonoids
and other different compounds such as saponin, tannin,
triterpenes and alkaloids (table1). With respect lipid
lowering capacity of this plant extract, it may be proposed
that the constituents of the plant extracts may act as
inhibitors for enzyme such as hydroxy-methyl-glutarylCoA reductase, which participates in de novo cholesterol
biosynthesis(table 4) as has been suggested for some

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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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