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International Journal of Research in Biological Sciences

Universal Research Publications. All rights reserved

ISSN 22499687
Original Article
ANTIDIABETIC ACTIVITY OF CLERODENDRUM PHLOMIDIS AGAINST
STREPTOZOTOCIN (STZ) INDUCED DIABETICS IN RATS
V. Lakshmi1 and G. Viji Stella Bai2
1

Research Scholar, Department of Botany, Kundavai Naachiyaar Government Arts College For Women (Autonomous),
Thanjavur, Tamil Nadu, S. India
2
Department of Botany, Kundavai Naachiyaar Government Arts College for Women (Autonomous),
Thanjavur, Tamil Nadu, S. India
Received XXXX 2015; accepted XXXX 2015

Abstract
The investigation was carried out to study the effects of Clerodendrum phlomidis leaves extract on blood glucose and lipid
profile level. The antidiabetic efficacy of the ethanolic extract of the leaves was evaluated in normal and Streptozotocin
(STZ) induced diabetic rats. The extract exhibited significant hypoglycemic and hypolipidemic activity in animal model
when compared with control group. The activity was also comparable to that of the effect produced by standard
antidiabetic agent Glibenclamide. The hypoglycemic produced by the extract may be due to increased uptake og glucose at
tissue level or increase in pancreatic betal cell function or due to inhibition of intestinal absorption of glucose. The study
indicated that the ethanolic extract is a potential antidiabetic agent and lends scientific support for its elses in folk
medicine.
2015 Universal Research Publications. All rights reserved
Keywords-: Glucose, Clerodendrum phlomidis, Streptozotocin, Lipid profile.
in Ayurvedic medicine system as well as in ethnomedicinal
1. Introduction
Diabetes is a metabolic disorder of carbohydrate, practices (Pareek et al., 2009). In India, indigenous
fat and protein, affecting a large number of population in remedies have been used in the treatment of diabetes
the world (Pareek et al., 2009). Diabetes mellitus is not a mellitus since the time of Charaka and Shusrutha. From the
single disorder but it is a group of metabolic disorder ethnobotanical information, about 800 plants which may
characterised by chronic hyperglycemia, resulting from possess anti-diabetic potential have been found (Warjeet
defects in insulin secretion, insulin action, or both. Singh, 2011; Patel et al., 2011). In the present study to
Increased thirst, increased urinary output, ketonemia and investigate the antidiabetic activity of Clerodendrum
ketonuria are the common symptoms of diabetes mellitus, phlomidis leaves on Streptozotocin (STZ) induced diabetic
which occur due to the abnormalities in carbohydrate, fat, rats.
and protein metabolism. When ketones body is present in 2. Materials and Methods
the blood or urine, it is called ketoacidosis, hence proper 2.1 Animals
treatment should be taken immediately, else it can leads to
Male albino rats of Wistar strain approximately
other diabetic complications (Craig et al., 2009). Diabetes weighing 160-180g were used in this study. They were
mellitus has caused significant morbidity and mortality due healthy animals purchased from the Indian Institute of
to
microvascular
(retinopathy,
neuropathy,
and Science, Bangalore. The animals were housed in spacious
nephropathy) and macrovascular (heart attack, stroke and polypropylene cages bedded with rice husk. The animal
peripheral vascular disease) complications (Thevenod, room was well ventilated and maintained under standard
2008). Diabetes is mainly attributed to the rapid rise in experimental conditions (Temperature 27 2 C and 12
unhealthy life style, urbanization and aging.
hour light/dark cycle) throughout the experimental period.
Herbal medications have been used for the All the animals were fed with standard pellet diet and water
treatment of variety of ailments, a huge number of were provided ad libitum. They were acclimatized to the
population in the world is entirely dependent on traditional environment for one week prior to experimental use. The
medicines (Feshani et al., 2011). A number of medicinal animal feed composition is crude protein (22.3%), crude oil
plants and their formulations areused for treating diabetes (4.01%), crude fibre (4.02%), Ash (8.02%) and sand silical
International Journal of Research in Biological Sciences XXXX; X(Y): ZZ

Table 1: Effect of Clerodendrum phlomidis on glucose and lipid profile in experimental rats
S.NO
Parameters
Group I
Group II
Group III
1.
Glucose (mg/dl)
84.74 5.93
347.45 28.32*
86.61 6.76
2.
Hb
17.90 1.25
8.24 0.57*
15.62 1.09
3.
Triglyceride (mg/dl)
77.77 5.44
252.77 17.69*
133.33 9.33
4.
Cholesterol (mg/dl)
36.36 2.54
136.36 9.54*
57.57 4.02
5.
HDL Cholesterol (mg/dl)
39.58 2.76
18.75 1.31*
29.16 2.04
6.
VLDL Cholesterol (mg/dl)
15.55 1.08
50.55 3.53*
26.66 1.86
7.
LDL Cholesterol (mg/dl)
12.33 0.86
104.56 7.31*
55.07 3.85

Group IV
103.38 7.23
14.20 0.99
138.88 9.72
63.63 4.45
29.16 2.04
27.77 1.94
62.24 4.35

Values are expresses as Mean SD for six rats

Significantly different from group I, III and IV (p<0.05)

(1.02%).
2.2 Chemicals:
Streptozotocin (STZ), Ethylene Diamine Tetra
Acetic Acid (EDTA)), Glibenclamide (Prudence Pharma
Chem, India), Chloroform were purchased for Sigma
chemical company, Mumbai All other chemicals and
reagents used in this study was of analytical grade with
high purity and were obtained from Glaxo laboratories and
Sisco Research laboratories, Mumbai, India.
2.3 Plant materials and preparation of extract
The fully mature Clerodendrum phlomidis leaves
were collected in April 2013 from Poyyundar kottai,
Thanjavur District, Tamil Nadu, India from a single herb.
The leaves were identified and authenticated by Botanist,
Dr. S John Britto, Department of Botany, St. Josephs
College, Tiruchirappalli, Tamil Nadu, India. A Voucher
specimen (VL 0.05) has been deposited at the Rapinat
Herbarium, St. Joseph,s College, Tiruchirappalli, Tamil
Nadu, India. The leaves were dried at room temperature
and coarsely powdered. The powder was extracted with
70% ethanol for 48 hours. A semi solid extract was
obtained after complete elimination of alcohol under
reduced pressure. The extract was stored in refrigerator
until used.
2.4 Streptozotocin (STZ) Induced Diabetic rats
Diabetes was induced in all groups except vehicle
control following overnight fasting(deprived of food for16h
allowed free access to water) by a single intraperitoneal
injection of 65mg/kg of streptozotocin (STZ) dissolved in a
freshly prepared 0.1M citrate buffer (pH4.5) (Liu et al.,
2008).The animals of vehicle control (Group I) were
injected with buffer alone. Diabetes was confirmed 72 h
after induction by measurement of tail vein blood glucose
levels by glucose oxidase-peroxidase method using strips.
Plant extracts at a dose of 500mg/kg was orally given once
a day for 15 days after hyperglycemia was confirmed by
the elevated glucose levels in blood determined at 72 h.
Glibenclamide is used as a standard at dose of 0.25mg/kg
(Arulmozhi et al., 2010).
2.5 Experimental Design
The animals were divided into four groups of six animals
each as follows:
Group -Vehicle control were injected with buffer alone
(Non-diabetic)
Group II -Diabetic control
Group III-Clerodendrum
phlomidis leaves extract
500mg/kg was orally given once a day for 21 days.

Group IV- Diabetic standard as 0.25mg/kg of

Glibenclamide, p.o was orally given once a day for 21
days.
2.6 Collection of blood and preparation of plasma
sample
At the end of the experimental period, the animals
were anaesthetized using chloroform vapour prior to
dissection. The blood was collected with EDTA as
anticoagulant. The blood was allowed to clot by standing at
room temperature for 30 minutes. The blood was
centrifuged at 3000rpm for 10minutes, and then the plasma
(supernatant) was isolated and stored at refrigerated until
required for analysis.
2.7 Biochemical estimations
Glucose was estimated by GOD/POD method
(Trinder, 1969). Cholesterol was estimated by Allain et al
(1974). Triglyceride was determined by the method of
Werner et al (1981). HDL cholesterol was estimated by the
method of Allain et al (1974). LDL VLDL cholesterol was
calculated as per Friedewalds (1972) equation.
Haemoglobin was estimated by Cyanmethaemoglobin
method (Dacie and Lewis, 1968) (Beacon Diagnostic Kit).
2.8 Statistical Analysis
Values were expressed as mean SD for six rats
in the each group and statistical significant differences
between mean values were determined by one way analysis
of variance (ANOVA) followed by the Tukeys test for
multiple comparisons (Harvey and Paige, 1998). The
results were statistically analyzed by Graphpad Instat
Software (Graphpad Software, San Diego, CA, USA)
version 3 was used and p< 0.05 was considered to be
significant.
3. Results
The ethanolic extract of of Clerodendrum
phlomidis was administered orally in an aqueous solution at
a dose of 500mg/kg body wt. to diabetic rats to assess the
synergetic impact of the plant extract. The plant extracts
were fed with normal and diabetes induced rats. The blood
glucose levels was significantly (P<0.05) reduced when
compared to the specific diabetic control animals.
The lipid profile such as TC, TG, LDL and VLDL
levels were significantly increased in diabetic control
animals (DC) whereas HDL levels were decreased when
compared to the control rats. The plant extract was
administered orally at a dose of 500mg/kg body wt., to
diabetic rats significant (P<0.05) depletion in the total
cholesterol, TG, LDL, and VLDL levels and increment of

International Journal of Research in Biological Sciences XXXX; X(Y): ZZ

Figure 1: shows the % of Antidiabetic activity of Clerodendrum phlomidis in experimental rats

Table 2 - % of Antidiabetic activity of Clerodendrum phlomidis in experimental rats
Groups
% of Antidiabetic activity
Group I
-Group II
-Group III
75.07
Group IV
70.24
HDL levels were recorded in the diabetic animals (Table
1).
4. Discussion
Insulin is the dominant hormone which
influencing the regulation of glucose metabolism. One of
the major effects of insulin is to enhance overall glucose
disposal by stimulation of glucose uptake into the target
tissues (Tiwari and Madhusudana, 2002). In diabetes the
increased blood sugar levels might be due to either insulin
resistance of the body cells or decreased secretion of
insulin from beta cells manifest in the decreased serum
insulin levels (Mohammad Ali, et al., 2004). Streptozotocin
(STZ) (2 deoxy -2- (3-methyl-3-nitrosuuredio) -D- gluco-pyranose) is commonly used for experimental induction of
type-I diabetes mellitus, which causes selective pancreatic
islet -cell cytotoxicity mediated through the release of
nitric oxide (NO).This results in rapid reduction in
pancreatic islet pyridine nucleotide concentration and
subsequent -cell necrosis. The action of STZ on
mitochondria generates SOD anions, which leads to
diabetic complications (Papaccio et al., 2000). Based on the
above perspectives, in the present study, the antidiabetic
activity has been assessed in rats made diabetic by STZ.
Sulfonylureas such as glibenclamide are often used as a
standard antidiabetic drug in STZ-induced diabetes to
compare the efficacy of variety of antihyperglycemic
compounds (Anderson et al., 1974).
The STZ selectively destroys the pancreatic cells
and induce hyperglycemia (Gilman et el., 1990, Kurup and
Bhonde., 2000, Jarvenin, 1995). The reduction in the serum
insulin levels in the STZ treated rats might be attributed to
the reduced secretion of the hormone which might be due
to the damage of the beta cells of endocrine pancreas were
recorded earlier studies (Yoon and Ray, 1985). Nitric oxide
has been demonstrated to participate in the beta cell

damage during STZ induced diabetes (Duran Reges et al.,

2004). Administration of Clerodendrum
phlomidis to
diabetic rats restored the levels of glucose. Present finding
is in agreement with Subramaniam et al. (2012) studies.
The insulin deficiency depletes the activity level
of lipoprotein lipase, thus leading to deranged lipoprotein
metabolism during diabetes (Ranganathan et al., 2000).
Diabetes affects both glucose and lipid metabolism
(Sperling et al., 2000). In the post prandial state elevated
serum insulin increases lipoprotein lipase activity in
adipose tissue and promotes fuel storage as triglycerides in
normal metabolism (Bhagavan, 2002).
The elevated hypertriglyceridemia was increased
in the synthesis of triglyceride rich lipoprotein particles
(very low density lipoprotein, VLDL) in liver diminished
catabolism in diabetic rats reported by Ginsberg (1991).
The lipoprotein levels in the STZ induced diabetic rats of
the present study reveal a significant alter in lipoprotein
metabolism. The serum total cholesterol content increased
significantly in diabetic animals. Since insulin has a potent
inhibitory effect on lipolysis in adipocytes, insulin
deficiency is associated with excess lipolysis and increased
influx of free fatty acids to the liver (Coppack, 1994, Ohno,
2000 ) The increased levels of low-density lipoprotein
(LDL) and very low density lipoprotein (VLDL) in the
diabetic animals might be due to over production of LDL
and VLDL by the liver due to the stimulation of hepatic
triglyceride synthesis as a result of free fatty acid influx
(Coppack, 1994). The high density lipoprotein (HDL) was
significantly reduced in the diabetic rats which indicate a
positive risk factor for atherosclerosis (Bopanna, 1997).
Supplementations of Clerodendrum phlomidis leaf extract
to diabetic rats restored the lipid profile. Our results
concord with the earlier work done by Kesari et al. (2006),
where it has been reported that lipid profile level in the

International Journal of Research in Biological Sciences XXXX; X(Y): ZZ

plasma is restored with the treatment of Aegle marmelos

seed extract in diabetic rats. After the administration of the
extract of Clerodendrum phlomidis to the STZ induced
diabetic rats revealed augmented serum insulin levels. The
increment of serum insulin levels might be due to increased
secretion of the hormone, which might reflect the probable
repair of the damaged beta cells of the endocrine of the
pancreas due to STZ.
The levels of serum TC, TG, LDL, and VLDL
were found to be significantly reduced in the plant extracts
treated diabetic animals. The blood glucose level of
Clerodendrum
phlomidis extract fed animal was
significantly (P<0.05) reduced. This might be due to the
reduced hepatic triglyceride synthesis and or reduced
lipolysis that might be due to the increase in serum insulin
levels in the plant extract treated rats. The HDL increased
significantly in the plant extract treated rats indicating a
reversed atherogenic risk.
Glycosylated haemoglobin (HbA1) was found to
increase in patients with diabetes mellitus to approximately
16% (Koenig et al., 1976) and the amount of increase is
directly proportional to the blood glucose level (Jackson et
al., 1979). In uncontrolled or poorly controlled diabetes
there is an increased glycosylation of a number of proteins
including haemoglobin and -crystalline of lens (Alberti
and Press, 1982). During diabetes the excess glucose
present in blood reacts with haemoglobin. Therefore, the
total haemoglobin level is decreased in STZ diabetic rats
The present study demonstrated that the
Clerodendrum phlomidis extract possess hypoglycemic
effect revealed by decreased serum lipid levels, restored
glucose and heamoglobin and therefore attributes to
therapeutic value of Clerodendrum
phlomidis leaves
extract to combat the diabetic condition in rats. The
potential antidiabetic activity of Clerodendrum phlomidis
extract due to the phytochemicals.
5. Acknowledgement
The authors are grateful to Dr. S. Velavan,
Director, Harman Institute of Science Education and
Research (www.harmanresearchcentre.com), Thanjavur,
Tamil Nadu for his support to prepare the manuscript.
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Source of support: Nil; Conflict of interest: None declared

International Journal of Research in Biological Sciences XXXX; X(Y): ZZ