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Bioscience Discovery, 8(3): 528-532, July - 2017
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Research Article
Ameliorative Effect of Aloe vera Leaf (Methanol) Extract on Blood
Glucose Level and Histopathology of Pancreas in Alloxan Induced
Diabetic Male Mice
Kadam J S and R N Patil
[email protected]
P.G. Department of Zoology, Sadguru Gadage Maharaj College,
Karad 415110, Maharashtra, India.
Article Info
Abstract
Received: 21-05-2017,
Revised: 25-06-2017,
Accepted: 27-06-2017
Diabetes mellitus is a chronic disease which is common throughout the world.
Apart from currently available synthetic drugs for diabetes like oral
hypoglycemic agents have side effects. Now days, many herbal medicines have
been recommended for the treatment of diabetes.The present study was designed
to examine the potential of Aloe vera leaf extract in alloxan induced diabetic
male mice. For investigation blood glucose level and histopathology of pancreas
tissue done. After administration of A. vera leaf extract (300mg/kg body weight).
The histopathology of the pancreatic islets and acini which nearly reverted to
their normal structure. The most important finding is that the blood glucose level
which once dropped to normal levels it is not elevated when extract feeding was
discontinued after 15 days.
Keywords:
Diabetes mellitus, Aloe
vera, hypoglycemic,
histopathology
INTRODUCTION
Diabetes mellitus is a chronic metabolic disorder
which is characterized by hyperglycemia and
glucose intolerance as a result of insulin deficiency
or intolerance of insulin action (Arika et al.,
2015).The metabolic abnormalities result in part
from a deficiency of blood glucose lowering
hormone insulin (Chandra et al., 2004). The
prevalence of diabetes is rising all over the world
due to population growth, aging, urbanization and
increase of obesity and physical inactivity. It is
assumed that the number of diabetic patients will
increase to 439 milion in 2030 which was 285
milion in 2010 (Shaw et al., 2010).
Pharmacological treatment of diabetes
mellitus is based on oral hypoglycemic agents and
insulin injections which have so many side effects,
also it has high cost which is not affordable in poor
economic communities. It is necessary to develop
traditional and alternative medicine (Arulrayan et
al., 2007). Herbal drugs widely used to treat a wide
variety of disease in present scenario. Indian folk
medicine comprises hundreds of herbal drugs for
therapeutic purposes (Syed Sabiha V, 2015).
Aloe vera is the oldest medicinal plant and most
applied medicinal plant worldwide (Rajeswari et
al., 2012). The plant leaves contains numerous
minerals, vitamins, enzymes, amino acids, natural
sugars and other bioactive compounds (Pankaj et
al., 2013). Aloe has been marketed as a remedy for
cough, wounds, ulcers, gastritis, headaches,
arthritis. Aloe vera is traditional remedy for tumor,
ulcer, viral activity (Chatterjee et al., 2013). The
present work was undertaken to study effect of
Aloe vera on blood glucose level and
histopathology of pancreas.
MATERIALS AND METHODS
Preparation of Aloe vera leaf extract
The fresh A.vera leaves (Voucher specimen: KJS-1)
were washed thoroughly with water, peel was
removed and only pulp was collected. The collected
pulp was lyophilized.
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Kadam and Patil
Extraction of lyophilized material was carried out
by soxhlet method (Aswar et al., 2011). The extract
obtained was dried at 370 C in oven. The obtained
yield was stored in refrigerator at 40C until further
use. The residual extract was resuspended in
distilled water and used in study as per desired
concentration when needed.
Animals
Healthy male mice (Mus musculus Linn.) were
used for present investigation. Breeding pairs were
obtained from (Appasaheb Birnale College of
Pharmacy, Sangali). Adult male mice of 5-6
months age and weighing about 28 to 30 ± 2 gm
were selected. All the animals were maintained
under standard laboratory conditions, with 12 hr
light and 12 hr dark cycle at temperature of 260 C ±
20 C in departmental animal house. The guidelines
of CPCSE were followed throughout the
experimentation. The animals were housed in
aluminium cages having dimensions of 10” x 8” x
5” and allowed to live in groups of 3-4 animals per
cage. They were fed with Amrut Mice feed,
marketed from Pranav Agro Industries, Pvt. Ltd.
Sangli and water ad libitum. The record of their
body weight was maintained.
Experimental design
Five-six month male mice were divided into three
groups containing 6 animals per group.
Control group: Mice were given intraperitoneal
injection of 0.15 M Acetate buffer PH 5.4 for 15
days.
Diabetic group: Mice were given single
intraperitoneal injection of alloxan 120 mg/kg body
weight (Fayed et al.,1988; Helal,2000 and Syiem et
al.,2002).
Recovery group: Mice from diabetic group were
given intraperitoneal injection of A. vera leaf
extract at dose of 300mg/kg body weight once a day
for 15 days (Rjasekaral et al.,2004).
Determination of blood glucose level
After the treatment fasting blood glucose was
measured by collecting a drop of blood from the tail
after incision with a sharp blade. The blood glucose
was determined by using a rapid glucose analyzer
with a glucose strip inserted in sugar scan digital
blood glucose monitoring glucometer. The result
were expressed in terms of milligram per deciliter
of blood (Kumar et al.,2006).
Histopathological study
After the completion of dose mice from all
experimental groups were killed by cervical
dislocation, pancreas were dissected out quickly and
fixed in 2% CAF fixative. Tissue were processed
and embedded in paraffin wax. Sections were cut at
5µ thickness and stained with Haematoxylene-eosin
and observed under light microscope. (Kleiner et
al.,2005).
Statistical analysis
The data was statistically analyzed by one way
ANOVA followed by Tukey HSD test.All the
values were expressed as mean ± SE. The
difference was considered significant when p ˂
0.001.
RESULTS AND DISCUSSION
Blood glucose level
The following table shows the level of blood
glucose in mice from control, diabetic and recovery
group. The mean level of glucose in the control
group was evaluated to be 81.8 ± 3.36 mg/dl (1:2, p
˂ 0.01whereas it was 227.4 ± 3.62 mg/dl in diabetic
group. After the treatment of diabetic mice with
A.vera leaf extract (300 mg/kg body weight) the
glucose level was found significantly decreased to
130.6 ± 3.45 mg/dl (2:3, p ˂ 0.01). The histological
structure of pancreas from control, diabetic and
treated group of mice observed under light
microscope is shown in microphotographs (Fig.1-
6).
Exocrine pancreas: The exocrine pancreas consists
of closely packed acini with connective tissue in
between them. The acinar cells were cuboidal in
shape, having small acinar lumen. The cells were
with rounded nuclei with dispersed and prominent
nucleoli (Fig. 1)
Table1: showing blood glucose level (mg/dl) in control and experimental mice.
Sr. No Animal group (n=6) Blood glucose level Statistical significance
1. Control 81.8 ± 3.36 1:2, P ˂ 0.01
2:3, P ˂ 0.01 2. Diabetic 227.4 ± 3.62
3. Recovery 130.6 ± 3.45
Values are mean ± S.E. (Numbers in parenthesis denotes number of animals)
p ˂ 0.01 =significant, p ˂ 0.001 = highly significant
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Histopathological study
In diabetic group disorganization of the structure of
exocrine cells with damaged and necrotic pancreatic
acini. Most acinar cells showed many small
vacuoles(Fig.2).
In recovery group the light
microscopographic observation showed that the
pancreatic acini had retained their cuboidal shaped
appearance as compared to diabetic group (Fig.3).
Endocrine pancreas: In control group microscopic
observation of islets of Langerhans showed cells
with large, rounded and basophilic nuclei and
prominent nucleoli ( Fig.4).
In diabetic group there were degenerative features
in most cells of islets. Nuclei became packed
together in groups and cytoplasm had lost their
granules. Cells also showed reduction in diameter
of islets of Langerhans as compared to control
group ( Fig.5).
In recovery group there was marked
improvement in islets of Langerhans and
regeneration of cells which might be due to
preventing the death of β cell as compared to
diabetic group (Fig.6).
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Kadam and Patil
Alloxan is the most commonly used agent
for the induction of diabetes for experimental study
in animal model. Alloxan causes diabetes by rapid
depletion of β cells by DNA alkylation and
accumulation of cytotoxic free radicals which result
into initial inflammation which is followed by
infiltration of activated macrophages and
lymphocytes. It results into reduction in plasma
insulin concentration leading to stable
hyperglycemia (Szkudelski, 2001). Hyperglycemic
condition occurs after 72 hr of alloxan induction. In
alloxan induced diabetic mice increased level of
glucose (227 ± 3.62) as compared to the control
(81.8 ± 3.36) mice in the present investigation was
might be due to reduced level of insulin.
Treatment of A. vera leaf extract in alloxan
induced diabetic mice started reduction in glucose
level. Effect of A.vera leaf extract at 300mg/kg
body weight dose was found to be more effective.
Haque et al., 2014 also found the same result. The
presence of flavonoids, alkaloids, tannins, saponins
which have shown that plant extract might be
possess hypoglycemic activity (Ohadoma et al.,
2011). This result agree with another finding of
Adesokan et al., 2006, Chcko et al., 2008,
Mohamed, 2011; Matheka, 2012. The exact
mechanism of action of Aloe vera is unclear.
Findings of previous studies suggest that Aloe vera
has multiple mechanism of action involved in
reduction of elevated plasma glucose in diabetes.
Administration of Aloe vera leaf extract
significantly reduced FPG levels in diabetic mice
while such activity is not observed in normal mice
(Noor et al., 2008; Shinde et al., 2013).
Previous studies propose that a disruption of the
normal relationship between insulin sensitivity and
pancreatic β-cell function is crucial for the
pathogenesis (Virally et al., 2007). On the other
hand histopathological observation of diabetic
pancreas caused vacuolization and necrosis of β
cells showed change in the shape of cells, this leads
to damage of exocrine as well as endocrine part of
pancreas ( Kalia et al., 2004). In the current study
the results showed that microscopic examination of
the pancreas of diabetic group revealed necrosis and
atrophy of β-cells of islets of Langerhans. This
results are in accordance with the findings of other
studies (Qadori, 2011; Khattab et al., 2013). The
histopathological observations confirmed the
improvement in the current study. Aggarwal (2015)
stated that extracts from natural products can
protect b-cells by enhancing the islets function and
restoring β cell mass. In addition the proliferation
and preservation of β-cell mass and function will be
the better way to prevent diabetes (Sharma et al.,
2015).
The level of blood glucose which were
raised in alloxan induced diabetic mice can be
lowered by Aloe vera leaf extract. Histopathological
investigation of pancreatic sections of diabetic mice
treated with Aloe vera extract revealed
approximately normal structure of acinar cells and
islets of Langerhans compared with the diabetic
group.Thus, further studies on bioactive or fractions
shall be evaluated for control and management of
diabetes mellitus.
Acknowledgement
Authors are thankful to the Department of Zoology,
S.G.M. College, Karad for providing basic
necessary facilities for research work.
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How to Cite this Article:
Kadam JS and RN Patil, 2017 Ameliorative Effect of Aloe vera Leaf (Methanol) Extract on Blood
Glucose Level and Histopathology of Pancreas in Alloxan Induced Diabetic Male Mice. Bioscience
Discovery, 8(3):528-532.