antidiabetic activity of hibiscus syriacus on alloxan ...antidiabetic activity of hibiscus syriacus...
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VOLUME 4 | NUMBER 5 | OCT | 2014 |140
Available online at www.ijntps.org | ISSN: 2277 – 2782 INTERNATIONAL JOURNAL OF NOVEL TRENDS IN PHARMACEUTICAL SCIENCES
Antidiabetic activity of Hibiscus syriacus on alloxan induced
diabetic rats
Kanjeri Nirosha*1, M.Divya1, R. Himabindu1, N. Himaja1, M. Pooja1, Mohemmed Sadiq2
*1Sree Vidyanikethan College of Pharmacy, Sree Sainath Nagar, Tirupati, A.Ranganpet, Andhra Pradesh-517102. 2Krishna Teja Pharmacy College, Chadalawada Nagar, Renigunta Road, Renigunta Road, Tirupati – 517501.
INTRODUCTION
Diabetes mellitus is a group of metabolic diseases
characterized by elevated blood glucose levels. It is
a result of defects in insulin secretion, insulin action
or both. Non‑Insulin Dependent Diabetes Mellitus
(NIDDM) is a multifactorial disease characterised by
hyperglycemia and lipoprotein abnormalities. These
traits are hypoyhetized to damage cell membranes,
which results in excessive generation of reactive
oxygen species. NIDDM has also been associated
with an increased risk for developing premature
atherosclerosis due to an increase in Triglyceride
(TG) and Low Density Lipoproteins (LDL) and
decrease in High Density Lipoprotein (HDL) levels
[1]. Chronic hyperglycemia is associated with
microvascular and macrovascular complications that
can lead to visual impairment, blindness, kidney
disease, nerve damage, amputations, heart disease,
and stroke. In 1997 an estimated 4.5% of the US
population had diabetes. Direct and indirect health
care expenses were estimated at $98 billion [2].
Epidemiology
It is estimated that 366 million people had DM in
2011; by 2030 this would have risen to 552 million
[3] the number of people with type 2 DM is
increasing in every country with 80% of people with
DM living in low- and middle-income countries. DM
caused 4.6 million deaths in 2011. It is estimated
that 439 million people would have type 2 DM by
the year 2030 [4]. The incidence of type 2 DM varies
substantially from one geographical region to the
other as a result of environmental and lifestyle risk
factors [5].
MATERIALS AND METHOD
Plant material
The whole plant of Hibiscus syriacus was collected
in kanipakam, chittoor district. The plant material
was authentified by Dr.K.Madhava chetty,
Assistant professor, department of botany, S.V.
University, Tirupathi. The plaant material was dried
in shade at room temperature. Then dried material
was powdered with the help of mechanical grinder
and subjected to seiving for equal particles of
coarse powder with the help of seiver.
Preparation of extract
The powdered plant material was packed in to
Soxhlet apparatus and defatted with petroleum
ether (60 - 80°C). Marc left was further extracted
To whom correspondence should be addressed:
K. Nirosha
Email: [email protected]
Article Info
Article history
Received 05 Oct 2014
Revised 10 Oct 2014
Accepted 12Oct 2014
Available online 30Oct 2014
Keywords
Diabetes Mellitus, Hibiscus
Syriacus, Blood Glucose.
Abstract
Diabetes mellitus is characterized by elevated blood glucose levels. The present
study was to evaluate the antidiabetic potential of Hibiscus syriacus on alloxan
induced diabetic rats. Animals are divided into five groups. For group I was
administered with distilled water (10 ml/kg/p.o), group II with alloxan
(120mg/kg/i.p), group III administered with alloxan and metformin (150mg/kg/p.o),
group IV was received alloxan and ethanolic extract (200 mg/kg/p.o), group V
received alloxan and ethanolic extract (400mg/kg/p.o). Administration of test and
standard drug for the period of 28 days, on 28th day biochemical parameters were
estimated. Results shows the decreased blood glucose level. Antidiabetic potential
was high in group V compared to group IV. Hence It concluded that high dose of
ethanolic extract shows the more antidiabetic potential compared to low dose.
RESEARCH ARTICLE
K. Nirosha et al., Antidiabetic activity of Hibiscus syriacus on alloxan induced diabetic rats
VOLUME 4 | NUMBER 5 | OCT | 2014 |141
with ethanol. Extract was air dried until it comes to
semi-solid to solid mass.
Phytochemical screening
The extract obtained was subjected to qualitative
tests for identification of different constituents like
tannins, alkaloids, saponins, glycosides, terpenes,
phenolics, flavonoids, carbohydrates, proteins and
steroids, by using simple and standard qualitative
methods described by Trease and Evans [6].
PHARMACOLOGICAL STUDY
Animals
Healthy albino rats of either sex weighing about
120-150 g were used during the study. The animals
were procured from Sree Vidyanikethan College of
Pharmacy, Tirupati, India. Before initiation of
experiment, the rats were acclimatized for a period
of 7 days. Standard environmental conditions such
as temperature ranging from 18 to 320C, relative
humidity ( 70%) and 12 hrs dark/light cycle were
maintained in the quarantine. All the animals were
fed with rodent pellet diet and water under strict
hygienic conditions. All procedures were performed
in accordance to CPCSEA guidelines after approval
from Sree Vidyanikethan College of Pharmacy,
Tirupati, India [Approval no. SVCP/IAEC/I-010/2013-
2014, dated on 14 march 2014.].
Acute oral toxicity study
The acute toxicity of ethanolic extract of Hibiscus
syriacus was carried out as per OECD-423 guidelines
for safe dose administration to animals. . The
animals were kept fasting for overnight providing
only water, after which the extracts were
administered orally at the dose level 2000mg/kg
body weight and observed for 14 days. the results
showed no sign of toxicity till 14 days at a starting
dose of 2000 mg/kg body weight. Hence, 1/10th
were considered as test dose for the study.
Induction of diabetes
Alloxan monohydrate was obtained from BROS
scientifics, tirupathi. To induce diabetes, alloxan 120
mg/kg, intraperitoneal (i.p.) Single dose will be
administered intraperitonial to groups II, III,IV&V.
The fasting blood glucose was determined after 72
hours7 Rats showing a blood glucose level of >200
mg/dl were taken for the study.
Experimental design
Rats having blood glucose level more than
200mg/dl taken for the study. Then rats are divided
into 5 groups, each group having 6 animals. Group
I: received normal saline (10ml/kg/day/p.o)
Group II: received alloxan (120mg/kg/i.p), Group III:
received standard drug Metformin (150mg/kg/p.o)
+ alloxan (120mg/kg/i.p), Group IV: received
ethanolic extract of Hibiscus syriacus
(200mg/kg/p.o) + alloxan (120mg/kg/i.p.), Group V:
received ethanolic extract of Hibiscus syriacus
(400mg/kg/p.o) + alloxan (120mg/kg/i.p.).
The study performed for 28 days.
Sample collection
28th day animals were sacrificed, pancreas was
isolated and it were fixed in 10% neutral formalin
solution. Fine sections were mounted on glass slides
and stained with hematoxylin Eosin for light
microscopic analysis.
Blood glucose estimation
Determination of blood glucose Fasting blood
glucose was estimated by commercially available
glucose strips (Accu- Chek) using One Touch
Glucometer (Johnson – Johnson, India). Estimation
of blood glucose level on 0th day, 7th day, 14th and
28thday.
RESULTS
Phytochemical screening: phytochemical screening of ethanolic extract was showed the presence of flavonoids,
tannins, saponins, glycosides and phenols. Carbohydrates, proteins, aminoacids and alkaloids are absent.
Table 1. Estimation of blood glucose level(mg/dl)
GROUPS 0TH DAY 7
TH DAY 14
TH DAY 28
TH DAY
Control 99.17±0.87 100±0.91 99.8±0.87 99.33±0.76
Diabetic control 230±2.9*a 230.5±2.91*a 235±1.68*a 239.7±0.98*a
Standard 230±3.1**b 157.7±1.9**b 116±1.40**b 113.2±0.90**b
EEHS(200mg/kg) 232±3.1*b 176±1.24*b 158±1.16*b 147.5±0.76*b
EEHS(400mg/kg) 232±3.1**b 162±1.72**b 129±2.62**b 128.2±o.70**b
K. Nirosha et al., Antidiabetic activity of Hibiscus syriacus on alloxan induced diabetic rats
VOLUME 4 | NUMBER 5 | OCT | 2014 |142
Values are expressed in ±SEM values (n=6). One‑way ANOVA followed by Dunnet’s multiple comparison tests.
Alloxan (120mg/kg.) was injected to except control , all other drug groups; a alloxan induced diabetic group vs
normal group, *p <0.001; b extract treated group vs alloxan induced diabetic group, *p <0.01; **p<0.001.
Fig 1. Estimation of blood glucose level
Fig 2. Histopathology of pancreas
A B
C
K. Nirosha et al., Antidiabetic activity of Hibiscus syriacus on alloxan induced diabetic rats
VOLUME 4 | NUMBER 5 | OCT | 2014 |143
D E
A – control- healthy β cells , B-Negative control- destruction of β cells, C-standard-recovery of β cells, D-
EEHS(200mg/kg)-slow recovery of β cells, E- EEHS(400mg/kg)-recovery of β cells better than “D”.
DISCUSSION
Pancreas is the primary organ involved in sensing
the organism’s dietary and energetic states via
glucose concentration in the blood and in response
to elevated blood glucose, insulin is Secreted.
Alloxan is one of the usual substances used for the
induction of diabetes mellitus apart from
streptozotocin by induce free radical production
and cause tissue injury. It has a destructive effect of
the beta cells of the pancreas [8]. A massive
reduction in insulin release by the destruction of
beta-cells of the islets of langerhans thereby
inducing hyperglycemia [9]. Alloxan has been
shown to Ethanolic Extract in the dose of 400
mg/kg reduced the reduction in the blood glucose
levels compared to Extract in the dose of 200
mg/kg. The literature reports reveal that flavonoids
and tannins present in the plant extract known to
possess antihyperglycemic activity.
Antihyperglycemic potential of test extract was
observed may be due to presence of similar
phytoconstituents which was evident by preliminary
phytochemical screening. In the present
investigation demonstrated ethanolic extract of
Hibiscus syriacus has the significant anti-diabetic
Activity.
CONLUSION
Present study was concluded that the ethanolic
extract of Hibiscus syriacus at the dose of 400 mg/kg
body weight produced significant antidiabetic
activity in alloxan-induced NIDDM in albino rats.
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