Ameliorative Effect of Aloe vera Leaf (Methanol) Extract

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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



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.

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Ameliorative Effect of Aloe vera Leaf (Methanol) Extract