“ANTIDIABETIC AND ANTIOXIDANT EFFECTS OF ETHANOLIC EXTRACT

OF Ficus carica LEAVES IN ALLOXAN INDUCED DIABETIC RAT”

Dissertation submitted to

THE TAMILNADU Dr. M.G.R MEDICAL UNIVERSITY

CHENNAI

In partial fulfillment of the requirements for the award of degree of

MASTER OF PHARMACY
in
PHARMACOLOGY

BY
Mr. JAYAKUMAR SP
(Reg. No.261226009)

Under the guidance of

MALINI SEN,M.Pharm.
Assistant Professor
Department of Pharmacology

MOHAMED SATHAK A.J. COLLEGE OF PHARMACY,

SHOLINGANALLUR, CHENNAI - 600119.

April-2014
 CERTIFICATE

This is to certify that the dissertation entitled “ANTIDIABETIC AND

ANTIOXIDANT EFFECTS OF ETHANOLIC EXTRACT OF Ficus carica

LEAVES IN ALLOXAN INDUCED DIABETIC RAT”submitted to The

Tamilnadu Dr. M.G.R. Medical university, Chennai, in partial fulfillment for the

award of degree of Master of Pharmacy in Pharmacology is a bonafide individual

research work done by Mr. JAYAKUMAR SP Mohamed Sathak A. J. College of

Pharmacy, Chennai, under the guidance and direct supervision of Mrs. MALINI

SEN during the academic year 2013-2014.

Place: Chennai

Date: (Dr. R. SUNDARARAJAN, M. Pharm., Ph.D)

Principal
Prof. M. Jagadeesan, M. Pharm
Professor and Head
Department of Pharmacology.

CERTIFICATE

This is to certify that the dissertation entitled “ANTIDIABETIC AND

ANTIOXIDANT EFFECTS OF ETHANOLIC EXTRACT OF Ficus carica

LEAVES IN ALLOXAN INDUCED DIABETIC RAT”submitted to The

Tamilnadu Dr. M.G.R. Medical university, Chennai, in partial fulfillment for the

award of degree of Master of Pharmacy in Pharmacology is a bonafide individual

research work done by Mr. JAYAKUMAR SP Mohamed Sathak A. J. College of

Pharmacy, Chennai, under the guidance and direct supervision of Mrs. MALINI

SEN during the academic year 2013-2014.

Place: Chennai

Date: (Prof. M. Jagadeesan, M.Pharm)

Head, Department of Pharmacology

MALINI SEN, M.Pharm
AssistantProfessor
Department of Pharmacology

CERTIFICATE

This is to certify that the dissertation entitled “ANTIDIABETIC AND

ANTIOXIDANT EFFECTS OF ETHANOLIC EXTRACT OF Ficus carica

LEAVES IN ALLOXAN INDUCED DIABETIC RAT”submitted to The

TamilnaduDr. M.G.R. Medical university, Chennai, in partial fulfillment for the

award of degree of Master of Pharmacy in Pharmacology is a bonafide individual

research work done by Mr. JAYAKUMAR SP Mohamed Sathak A. J. College of

Pharmacy, Chennai, under my guidance and direct supervisionduring the academic

year 2013-2014.

Place: Chennai

Date: (MALINI SEN, M.Pharm)

Guide and Supervisor

Mr. JAYAKUMAR SP
II year- M. Pharm, Pharmacology (Reg.no: 261226009),
Department of Pharmacology,
Mohamed Sathak A. J. College of Pharmacy.

DECLARATION OF THE CANDIDATE

I hereby declare that the thesis titled “ANTIDIABETIC AND

ANTIOXIDANT EFFECTS OF ETHANOLIC EXTRACT OF Ficus carica

LEAVES IN ALLOXAN INDUCED DIABETIC RAT”submitted in partial

fulfillment for the award of degree Master of Pharmacy to The TamilnaduDr.

M.G.R. Medical University and carried out at Mohamed Sathak A.J.College of

Pharmacy, Chennai, is my original and independent work done under the direct

supervision and guidance Mrs.MALINI SEN, M. Pharm, Associate Professor,

Department of Pharmacology during the academic year 2013-2014 and this thesis

contains no material which has been accepted for the award of any degree or

diploma of other Universities.

Place: Chennai

Date: [JAYAKUMAR SP]

 ACKNOWLEDGEMENT

I wish to express my gratitude towards “GOD – Almighty”, who gave me

the strength and courage to fulfill my dream and has showered upon me his choicest
blessings.

I take this opportunity to express my heartfelt thanks to all those, who

knowingly or unknowingly contributed to the success of my dissertation work.

My heartfelt thanks to my parents and my brother and sisters for their love
affection and constantly encouraging, guiding when I thought nothing is happening.

I wishtoexpressmy deepestgratitudeto Management of Mohamed

Sathak Trust, Chennai and Management of Mohamed Sathak A.J. College of
Pharmacy, in acknowledging all facilities provided to use at the institution enabling
us to do work of this magnitude.

I express my sincere thanks to Dr. R. Sundararajan, M.Pharm., Ph.D.,

Principal, Mohamed Sathak A.J. College of Pharmacy, for his moral encouragement
and providing necessary facilities required for my dissertation work.

It is indeed a great pleasure to express my deep sense of gratitude and

humble thanks to my guide Mrs.MALINI SEN, M. Pharm, Assistant Professor
Mr. J. Gunasekaran, M. Pharm, (Ph.D.), Associate Professor &
Prof. M. Jagadeesan, M. Pharm, Head of the Department of Pharmacology,
Mohamed Sathak A.J. College Of Pharmacy, Chennai, for his invaluable guidance
and constant encouragement that formed the foundation of this project. His
discipline, principle, simplicity, the profound knowledge and the subject
understanding influenced me a lot. I am proud to say that it has been a most fruitful
and enjoyable experience to work under his untiring and dynamic guidance.
 I am deeply indebited to the teaching staff especially Dr. Deepa Sankar,
M. Pharm., Ph.D., Vice Principal, Mohamed Sathak A.J. College of Pharmacy,
M. Komala, M. Pharm, (Ph.D), HOD and Mrs. N. B. SanthaSheela,
M. Pharm.,(Ph.D)., Associate Professor, Department of Pharmaceutics and other
teaching staff including Mr. S. Ramachandran, and Mr. Shakti Saravanan,
M. Pharm.,who was always a source of knowledge and inspiration to me and also for
their prompt assistance and cooperative attitude.

I thank Mr. A. Mohamad Jamaludeen, lab assistant, Department of

Pharmacology for his timely help.

I wish to express my special thanks to librarians Dr. M. Amudha,

M.A.L.I.Sc., Ph.D., and Mrs. Kumari, M.A.L.I.Sc., for helping me in collecting
my reference material.

I also wish to express my sincere thanks to Mr. Arunachalam and

Mr. Mani, for his technical support and advices given during the entire course of
my project work. With his dynamic approach he boosted my morale, which helped
me in completion of this dissertation.

Friends are integral part of life, so I take this opportunity to thank my

dearest friends Aleemsha, Mohamed Faisal, Padmapriya, Neelima Maddali,
Ananthula Prudhvidhar, Anandraj Kumar, Dinesh Kumar, Ramdoss, and
Saravanan who always pushed my confidence and creativity to the eventual extent
of my mind and for their unflinching support and co-operation during my
dissertation.

Also I want to thank all teaching and non teaching staff, who directly or
indirectly helped me in completing this dissertation work successfully.

JAYAKUMAR SP
 CONTENTS

S.No. TOPIC Page No.

I INTRODUCTION 1

PLANT PROFILE AND REVIEW OF

II
LITERATURE 11

III AIM AND OBJECTIVES 15

IV PLAN OF WORK 16

V MATERIALS AND METHODS 17

VI RESULTS AND DISCUSSION 25

VII SUMMARY AND CONCLUSION 36

VIII BIBLIOGRAPHY 37
 LIST OF TABLES

TABLE TITLE PAGE

NO NO.

1 Phytochemical screening results of Anjir leaves 27

2 Anti Diabetic Activity of Ethanolic Extract of Anjir 29

leaves on Alloxan Induced Diabetic Rats

3 Antioxidant Activity of Ethanolic Extract of Anjir 31

leaves on Alloxan Induced Diabetic Rats

4 Hepatic Glycogen Level of Ethanolic Extract of Anjir 32

leaves on Alloxan Induced Diabetic Rats

5 Serum Cholesterol Level of Ethanolic Extract of Anjir 33

leaves on Alloxan Induced Diabetic Rats

 LIST OF FIGURES

FIGURENO TITLE PAGE

NO.

1 Type 1 Diabetes: Insufficient Insulin 4

2 Information on Type 1 Diabetes 4

3 Factors Contributing To Type 2 Diabetes 6

4 Anjir plant 13

5 Plan of Work 18

6 Anti Diabetic Activity of Ethanolic Extract of Anjir 30

leaves

7 Antioxidant Activity of Ethanolic Extract of Anjir 31

leaves

8 Hepatic Glycogen Level of Ethanolic Extract of Anjir 32

leaves

9 Serum Cholesterol Level of Ethanolic Extract of Anjir 33

leaves
 ABBREVATIONS

µg - Microgram
AL - Anjir Leaves
ALB - Albumin
ANOVAs - Analysis of Variance
CAT - Catalase
GLO - Globulin
gm - Gram
I.U - International units
LD - Lethal Dosage
LPO - lipid peroxidase
mg - Milligram
mg/dl - Milligram per deciliter
mg/kg - Milligram per kilogram
SOD - Superoxide Dismutase
TP - Total Protein
U/L - Unit per Liter
 1. INTRODUCTION

1.1. Medicinal Plants

The therapeutic properties of medicinal plants are conditioned by the

presence of active substances, such as alkaloids, flavonoids, glycosides,
vitamins, tannins, and coumarin compounds, which physiologically affect
humans and animals or which are biologically active in relation to the causative
agents of various diseases. A special group of medicinal plants are antibiotics
[1].

Harvested medicinal plants are usually dried in special crop dryers, in

lofts, or in the shade. To obtain essential oils and juices from certain medicinal
plants only freshly harvested material can be used. Dried medicinal plants are used
in pharmaceutical practice, in galenics production, and in the chemical and
pharmaceutical industry. The composition and quantity of active substances found in
different organs of medicinal plants vary and change in the course of the year as a
result of the aging of the plant and habitat conditions [2]. Those parts of the plants in
which the largest quantity of these substances accumulates are collected first.

Many medicinal plants are no longer used, owing to the availability of

more effective drugs. However, more than 30 percent of all drugs obtained in the
USSR are of plant origin; for the most part they are less toxic than synthetic agents
and have no side effects. However, treatment with medicinal plants must be
conducted under the supervision of a physician. More than 30,000 tons of raw
materials from approximately 220 species of medicinal plants are used annually in
the USSR. Of the plants collected, more than 75 percent of the species grow wild,
accounting for 50 percent of the total weight. The rest are cultivated on 23
sovkhozes of the Ministry of Medicinal Industry. The opium poppy and peppermint
are also cultivated on kolkhozes. A number of medicinal plants are cultivated and
gathered in their natural habitats, including marshmallow, henbane, valerian,
ginseng, Saint-John‟s-wort, plantain, motherwort, and burmarigold. In gathering the
raw materials, only part of the plant should be dug up or cut off in order to ensure its
self-renewal. Only a few medicinal plants are imported into the USSR. A few dozen
species of medicinal plants are exported annually from the USSR, including several
thousand tons of licorice roots. Many medicinal plants are used in the food industry,
in the perfume industry and in metallurgy [3].

The All-Union Scientific Research Institute of Medicinal Plants, a number

of institutes of the Academy of Sciences of the USSR and of the Academies of
Sciences of the Soviet republics, pharmaceutical institutes (pharmaceutical
departments), botanical gardens, and other scientific research and educational
institutions are searching for new preparations of plant origin, cultivating medicinal
plants and studying their natural properties, and creating a rational regime for their
use [3].

1.2. Diabetes

Diabetes is a metabolic disorder that is characterized by high blood

glucose and either insufficient or ineffective insulin. 5.9% of the population in the
United States has diabetes, and diabetes is the seventh leading cause of death in
India. Diabetes is a chronic disease without a cure, however, with proper
management and treatment, diabetics can live a normal, healthy lives. There are two
main types of diabetes, Type I and Type II, described below [4].

Type I Diabetes

Insulin-dependant is caused by damage to the pancreas. The pancreas

contains beta cells, which make insulin. With Type I diabetes, the deficiency of
insulin is due to a decline in the number of beta cells the pancreas contains. It
appears that certain genes make Type I diabetics more susceptible, but a triggering
factor (usually a viral infection) sets it off. In most people with Type I diabetes, the
immune system makes a mistake, attacking the beta cells and causing them to die.
Without the beta cells, you cannot produce insulin. Glucose then builds up in the
blood and causes diabetes [5]. Type I diabetes exhibits the following warning signs.
 1. Losing weight without trying

2. An increased need to urinate

3. Increased hunger

4. Increased thirst

5. Trouble seeing

6. Feeling tired and/or

7. Going into a coma

For Type I diabetes, treatment usually consists of a healthy diet, exercise,

and insulin shots to replace the insulin that your body no longer produces. Most
insulin-dependent diabetics test their blood at least four times per day to monitor
their blood‟s glucose level. This is necessary to keep their blood glucose within
certain limits. If blood glucose is not monitored, and if insulin levels are not kept in
check, three things may happen:

Ketoacidosis – occurs when your blood glucose levels are highly elevated, by
either eating too much or taking too little insulin, by stress or illness. In this case,
there is too little insulin in the blood. Your body then begins breaking down fat for
energy, producing chemicals called ketones. Ketones can make you throw up, have
difficulty breathing, cause excessive thirst, cause dry, itchy skin, or even cause
coma.

Hypoglycemia – occurs when blood glucose levels become too low. It can be
by taking too much insulin, eating too little, skipping meals, eating at the wrong
time, exercising too intensely or for too long, or by drinking alcohol on an empty
stomach. If your blood glucose is too low you may feel hungry, confused, tired,
shaky or nervous [6].

Complications – elevated glucose levels in the blood over time can hurt your
organs. Diabetes can damage kidneys, eyes and nerves, and makes heart and blood
vessel disease more likely. Diabetics can defend themselves from complications by
keeping their glucose levels under control [7].

Figure 1: Type 1 Diabetes: Insufficient Insulin

Figure 2: Information on Type 1 Diabetes

 Type II Diabetes

Type II diabetes is the most common form of diabetes, with about 90% of
diabetes falling into the Type II category. With Type II diabetes, glucose builds up
in the blood – not because not enough insulin is present, but probably because cells
lose their insulin receptors and become less sensitive to insulin. Type II diabetes
usually (though not always) occurs in individuals who are over 40 years of age who
are overweight.

Type II diabetes produces mild symptoms, and can be controlled with a

healthy diet, exercise and weight loss. Type II diabetics should also monitor their
glucose levels to be sure they are maintaining healthy levels. In some cases, weight
loss, diet and exercise are not enough to control the glucose levels. In those cases,
your physician may control your diabetes by prescribing diabetes pills or insulin
shots. Type II diabetes can cause three types of problems.

1. High Blood Sugar – high glucose levels in the blood are most likely
when you‟re sick or under a lot of stress. High blood sugar can cause
you to have a headache, blurry vision, excessive thirst and an
increased need to urinate, and cause dry, itchy skin. Though less of a
problem with Type II diabetes, ketones can build up in the blood
when Type II diabetics have symptoms of high blood sugar, or when
they are sick.

2. Low Blood Sugar – When blood sugar falls to low you may feel tired,
shaky, nervous, hungry or confused. It may be caused by taking too
much diabetes medicine, eating too little or skipping meals,
exercising too intensely or for too long, or from drinking alcohol
without eating.

3. Complications – Elevated blood glucose over many years can hurt

organs, including the eyes, kidneys, and eyes. It can also make heart
and blood vessel disease more likely. The best defense against
complications is a careful monitoring of blood glucose, a healthy diet
and exercise [8].
 Figure 3: Factors Contributing To Type 2 Diabetes

Other Forms of Diabetes

Gestational diabetes occurs during pregnancy and affects about 4% of all

pregnant women, approximately 135,000 cases in the U.S. per year.

Gestational diabetes usually goes away after pregnancy, but once you've
had gestational diabetes, your chances are higher that it will happen in future
pregnancies. In some women pregnancy uncovers Type 1 or Type 2 diabetes and
these women will need to continue diabetes treatment after pregnancy.

There seems to be a link between the tendency to have gestational

diabetes and Type 2 diabetes, and many women who had gestational diabetes
develop Type 2 diabetes later on. Gestational diabetes and Type 2 diabetes both
involve insulin resistance. Certain basic lifestyle changes may help prevent diabetes
after gestational diabetes.
 Pre-diabetes is a condition that causes a person‟s blood sugar levels to be
higher than normal but not high enough to be diagnosed with diabetes. The
American Diabetes Association estimates that there are 41 million Americans that
have pre-diabetes in addition to the 18.2 million with diabetes.

Before people develop Type 2 diabetes, they almost always have

“pre-diabetes.” Recent research has shown that some long-term damage to the body,
especially the heart and circulatory system, may start during pre-diabetes.

Risks for Diabetes

1. Individuals with parents or siblings with diabetes

2. People over the age of 45

3. People who are overweight

4. People who do not exercise regularly

5. People with low HDL cholesterol or high triglycerides

6. Certain racial and ethnic groups (African Americans, Latinos, Asians

and Native Americans)

7. Women who had gestational diabetes or who had a baby weighing

9 pounds or more at birth.
 Antioxidant

An antioxidant is a substance that when present in low concentrations

relative to the oxidizable substrate significantly delays or reduces oxidation of the
substrate

Antioxidants get their name because they combat oxidation. They are
substances that protect other chemicals of the body from damaging oxidation
reactions by reacting with free radicals and other reactive oxygen species within the
body, hence hindering the process of oxidation. During this reaction the antioxidant
sacrifices itself by becoming oxidized. However, antioxidant supply is not unlimited
as one antioxidant molecule can only react with a single free radical. Therefore,
there is a constant need to replenish antioxidant resources, whether endogenously or
through supplementation [9].

Free Radical Formation

Atoms are most stable in the ground state. An atom is considered to be

“ground” when every electron in the outermost shell has a complimentary electron
that spins in the opposite direction. By definition a free radical is any atom with at
least one unpaired electron in the outermost shell, and is capable of independent
existence [10]. A free radical is easily formed when a covalent bond between entities
is broken and one electron remains with each newly formed atom. Free radicals are
highly reactive due to the presence of unpaired electron(s). The following literature
review addresses only radicals with an oxygen center. Any free radical involving
oxygen can be referred to as reactive oxygen species. Oxygen centered free radicals
contain two unpaired electrons in the outer shell. When free radicals steal an
electron from a surrounding compound or molecule a new free radical is formed in
its place. In turn the newly formed radical then looks to return to its ground state by
stealing electrons with antiparallel spins from cellular structures or molecules. Thus
the chain reaction continues and can be thousands of events long. The electron
transport chain (ETC), which is found in the inner mitochondrial membrane, utilizes
oxygen to generate energy in the form of adenosine triphosphate (ATP). Oxygen
acts as the terminal electron acceptor within the ETC. The literature suggests that
anywhere from 2 to 5% (14) of the total oxygen intake during both rest and exercise
have the ability to form the highly damaging superoxide radical via electron escape.
During exercise oxygen consumption increases 10 to 20 fold to 35-70 ml/kg/min. In
turn, electron escape from the ETC is further enhanced. Thus, when calculated, .6 to
3.5 ml/kg/min of the total oxygen intake during exercise have the ability to form free
radicals. Electrons appear to escape from the ETS at the ubiqunone-cytochrome c
level [11].

Peroxidation

Polyunsaturated fatty acids (PUFAs) are abundant in cellular membranes

and in low-density lipoproteins. The PUFAs allow for fluidity of cellular
membranes. A free radical prefers to steal electrons from the lipid membrane of a
cell, initiating a free radical attack on the cell known as lipid peroxidation. Reactive
oxygen species target the carbon-carbon double bond of polyunsaturated fatty acids.
The double bond on the carbon weakens the carbon-hydrogen bond allowing for
easy dissociation of the hydrogen by a free radical. A free radical will steal the
single electron from the hydrogen associated with the carbon at the double bond. In
turn this leaves the carbon with an unpaired electron and hence becomes a free
radical. In an effort to stabilize the carbon-centered free radical molecular
rearrangement occurs. The newly arranged molecule is called a conjugated diene.
The CD then very easily reacts with oxygen to form a proxy radical. The proxy
radical steals an electron from another lipid molecule in a process called
propagation. This process then continues in a chain reaction [12, 13].

Types of Free Radicals

There are numerous types of free radicals that can be formed within the
body. The most common ROS include: the superoxide anion, the hydroxyl radical,
singlet oxygen, and hydrogen peroxide Superoxide anions are formed when oxygen
acquires an additional electron, leaving the molecule with only one unpaired
electron. Within the mitochondria O2- is continuously being formed. The rate of
formation depends on the amount of oxygen flowing through the mitochondria at
any given time. Hydroxyl radicals are short-lived, but the most damaging radicals
within the body. These reactions are significant as the substrates are found within
the body and could easily interact. Hydrogen peroxide is produced in vivo by many
reactions. Hydrogen peroxide is unique in that it can be converted to the highly
damaging hydroxyl radical or be catalyzed and excreted harmlessly as water.
Glutathione peroxidase is essential for the conversion of glutathione to oxidized
glutathione, during which H2O2 is converted to water. If H2O2 is not converted into
water is formed. Singlet oxygen is not a free radical, but can be formed during
radical reactions and also cause further reactions. Singlet oxygen violates Hund's
rule of electron filling in that it has eight outer electrons existing in pairs leaving one
orbital of the same energy level empty. When oxygen is energetically excited one of
the electrons can jump to empty orbital creating unpaired electrons [14]. Singlet
oxygen can then transfer the energy to a new molecule and act as a catalyst for free
radical formation. The molecule can also interact with other molecules leading to the
formation of a new free radical.

Physiological Effects

Under normal conditions (at rest) the antioxidant defense system within
the body can easily handle free radicals that are produced. During times of increased
oxygen flux (i.e. exercise) free radical production may exceed that of removal
ultimately resulting in lipid peroxidation. Free radicals have been implicated as
playing a role in the etiology of cardiovascular disease, cancer, Alzheimer‟s disease,
and Parkinson‟s disease. While worthy of a discussion these conditions are not the
focus of the current literature review. This literature review will only examine the
current literature addressing the relationship between free radicals and exercise,
which is introduced below. The driving force behind these topics is lipid
peroxidation. By preventing or controlling lipid peroxidation the concomitant effects
discussed below would be better controlled.
 Requirement

Oxygen consumption greatly increases during exercise, which leads to

increased free radical production. The body counters the increase in free radical
production through the antioxidant defense system. When free radical production
exceeds clearance oxidative damage occurs. Free radicals formed during chronic
exercise may exceed the protective capacity of the antioxidant defense system,
thereby making the body more immune to disease and injury. Therefore, the need
for antioxidant supplementation is discussed.

Fatigue

A free radical attack on a membrane usually damages a cell to the point

that it must be removed by the immune system. If free radical formation and attack
are not controlled within the muscle during exercise a large quantity of muscle could
easily be damaged. Damaged muscle could in turn inhibit performance by the
induction of fatigue. The role individual antioxidants have in inhibiting this damage
has been addressed within the review of the four antioxidants that follows.

Recovery

One of the first steps in recovery from exercise induced muscle damage is
an acute inflammatory response at the site of muscle damage. Free radicals are
commonly associated with the inflammatory response and are hypothesized to be
greatest twenty-four hours after completion of a strenuous exercise session. If this
theory were valid then antioxidants would play a major role in helping prevent this
damage. However, if antioxidant defense systems are inadequate or not elevated
during the post-exercise infiltration period free radicals could further damage muscle
beyond that acquired during exercise. This in turn would increase the time needed to
recover from an exercise bout [15].

Importance of Free Radicals

This section has focused only on the negatives associated with free radical
production. However, free radicals are naturally produced by some systems within
the body and have beneficial effects that cannot be overlooked. The immune system
is the main body system that utilizes free radicals. Foreign invaders or damaged
tissue is marked with free radicals by the immune system. This allows for
determination of which tissue need to be removed from the body. Because of this
some question the need for antioxidant supplementation, as they believe
supplementation can actually decrease the effectiveness of the immune system.

Antioxidant Defenses

Antioxidant means “against oxidation.” Antioxidants work to protect

lipids from peroxidation by radicals. Antioxidants are effective because they are
willing to give up their own electrons to free radicals. When a free radical gains the
electron from an antioxidant it no longer needs to attack the cell and the chain
reaction of oxidation is broken [16]. After donating an electron an antioxidant
becomes a free radical by definition. Antioxidants in this state are not harmful
because they have the ability to accommodate the change in electrons without
becoming reactive. The human body has an elaborate antioxidant defense system [17].
 2. PLANT PROFILE AND REVIEW OF LITERATURE

2.1. Botanical classification

Scientific Name: Ficus carica

Other Names: Common Fig, Edible Fig, Anjir, Anjuru, Anjura, Athi, Seemai,
Simayathi, Figue (French), Feige (German), Figo (Italian & Portugese)

Taxonomy:

Kingdom : Plantae

Division : Magnoliophyta

Class : Magnolipsida

Order : Rosales

Family : Moraceae

Genus : Ficus

Species : F. carica

Ficus is a pan-tropical genus of trees, shrubs and vines occupying a wide

variety of ecological niches; most are evergreen, but some deciduous species are
endemic to areas outside of the tropics and to higher elevations [18]. Fig species are
characterized by their unique inflorescence and distinctive pollination syndrome,
which utilizes wasp species belonging to the Agaonidae family for pollination.

2.2. Macroscopic character

Ficus constituted one of the largest genera of medicinal plants with about
750 species of woody plants, trees, and shrubs primarily occurring in subtropical and
tropical regions throughout the world. The genus are remarkable for the large
variation in the habits of its species [19]. In India, the most important species of
Ficus are F. bengalensis, F. carica, Ficus racemosa and F. elastica. Ficus carica is
commonly referred as Fig. Various parts of the plant like bark, leaves, tender shoots,
fruits, seeds, and latex are medicinally important. The fig is a very nourishing food
and used in industrial products. It is rich in vitamins, mineral elements, water, and
fats. Figs are one of the highest plant sources of calcium and fiber. According to
USDA data for the Mission variety, dried figs are richest in fiber, copper,
manganese, magnesium, potassium, calcium, and vitamin K, relative to human
needs. They have smaller amounts of many other nutrients. Figs have a laxative
effect and contain many antioxidants. They are good source of flavonoids and
polyphenols [20] and some bioactive compounds such as arabinose, β-amyrins,
β-carotines, glycosides, sitosterol and xanthotoxol [21-23]. The dried figs produced
a significant increase in plasma antioxidant capacity and also used in various
disorders such as gastrointestinal respiratory, inflammatory, cardiovascular
disorders, ulcerative diseases, and cancers [24-27].

In traditional medicine the roots are used in treatment of leucoderma and

ringworms and its fruits which are sweet, have antipyretic, purgative, aphrodisiac
properties and have shown to be useful in inflammations and paralysis [28, 29].
F. carica has been reported to include antioxidant, antiviral, antibacterial,
hypoglycemic, cancer suppressive, hypotriglyceridaemic, and anthelmintic effects
[30-32]. This study was aimed to present an overview of traditional, phytochemical
and pharmacological investigations of bioactive compounds present in this plant.

Figure 4: Anjir plant

2.3. Medicinal Uses of Fig

1. Grind four fig leaves along with sugar candy. Regularly take this
mixture along with a glass of water two times in a day. This fig home
remedy is useful in the natural treatment of liver cirrhosis.

2. Consuming one teaspoon of fig seeds along with a teaspoon of honey

daily to control diabetes. Follow this therapy for about a week.

3. Fig benefits in removing kidney stones by having a cup of fresh fig

juice for a few days. In addition, you can take a solution of six figs
boiled in a cup of water. Follow this fig remedy for about a month
[33]

4. Take a cup of buttermilk and add a teaspoon of powdered fig bark in

it. Drink this solution daily for a few days to get rid of diarrhea.

5. When dealing with excessive menstruation, apply freshly ground ten

fig leaf buds on the lower abdomen. Leave it for a couple of hours
and repeat the procedure a few times.

6. Soak a couple of figs in milk overnight and consume them next

morning to increase sexual vitality.

7. Those suffering from anemia can eat 2-3 figs soaked in a cup of
water overnight. Consume these figs along with milk in the morning
for a month.

8. Take one and a half liters of water and add 50 g each of dried figs,
barley and raisins in it. Boil the solution. Simmer and keep it covered
for about 15 minutes. Next, add 15 g of chopped licorice root in it.

9. Cover and leave it to stand overnight. Finally, stain the liquid and
take one teaspoon of this solution. This is an effective figs home
remedy for constipation in children.

10. Applying the milky white juice of fig leaves on the affect areas cures
corns and calluses. It is useful in treating insect bites, too.

11. Chewing a couple of tender fig leaves and then rinsing the mouth
with warm water reduces bad breath and mouth ulcers [34].
2.4. Review of Literature

 Rahul Kumar Modi, et al, has reported a review on: Comparative

studies on ethanolic extract of root and stem bark of Ficus carica for
analgesic and anti-inflammatory activities [35].

 Deepika Paliwal, et al, was reported the Preliminary and

pharmacological profile of Ficus religiosa l [36].

 Vikas. V patil, et al, gave a over view on Ficus carica Linn, Research
Jouralof Medicial Plants [37].

 Baby Joseph and S.Justin Raj. Pharmacognostic and phytochemical

properties of Ficus carica Linn [38].

 K. K. Gangwar, et al, has reported the Ethnomedicinal Plant

Diversity in Kumaun Himalaya of Uttarakhand, India [39].

 G.B. Kavishankar et al, has reported the Diabetes and medicinal

plants-A review [40].

 Mohamed Bnouham et al, has reported the nature of Medicinal plants

with potential antidiabetic activity - A review of ten years of herbal
medicine research [41].

 P. A. Akah, et al reported the Antidiabetic activity of aqueous and

methanol extract and fractions of Gongronema latifolium
(Asclepidaceae) leaves in Alloxan Diabetic Rats [42].

 Mishra Shanti Bhusan et al reported the analytical review of plants

for anti-diabetic activity with their phytoconstituent & mechanism of
action [43].

 Kanika pandit, et al, has evaluate the anti hyperlipidemic activity of

Ficus hispida Linn leaves. International journal of pharmacy and
pharmaceutical sciencesvol-3, iss-5, 188-191, 2011 [44].
 AIM AND OBJECTIVES OF STUDY

AIM

To evaluate Antidiabetic and Antioxidant Effects of Ethanolic Extract of

Ficus carica Leaves In Alloxan Induced Diabetic Rat.

OBJECTIVES OF THE STUDY

The objectives of the present study is to

 Identify the phytochemical constituents of the medicinal plant Ficus

carica

 To determine the acute toxicity profile of plant Ficus carica.

 To authenticate antidiabetic effect of the medicinal plant through

standard evaluation techniques.

 To evaluate antioxidant property of Ficus carica

 3. PLAN OF WORK

SURVEY

PLANT SELECTION

PLANT
AUTHENTICATION

LITERATURE SURVEY

PROCESSING OF PLANT
MATERIAL

EXTRACTION ANTI-DIABETIC
AND
ANTI-OXIDANT
SCREENING
PRELIMINARY
PHYTOCHEMICAL SCREENING

Figure 5: Plan of work

 4. MATERIALS AND METHODS

4.1. Collection and authentication of Anjir leave

Leaves of Anjir leaves were collected in June 2013 from Sri

Venkateswara University, Tirupati, India. The leaves were verified and
Authenticated by Dr. K. Madhava Chetty, Assistant Professor, Department of
Botany, Tirupathi. The leaves were dried in shade and made into coarse powder.

4.2. Preparation of ethanolic extracts of Anjir leaves

The leaves were separated from the plant and it was washed with absolute
ethanol to avoid the microbial growth, the leaves were dried at open air under the
shade, cut in to small pieces and powdered mechanically, then 50 gm of powder
Anjir leaves was extracted with 250 ml ethanol in a soxhlet apparatus for 72 hrs. The
extract obtained was concentrated by recovery of ethanol. The concentrated product
was used as ethanolic extract of leaves of Anjir.

4.3. Phytochemical investigation on Anjir leaves [45, 46, 47]

4.3.1. Test for saponins

Foam test

Take 2 ml of drug solution in a test tube, add small amount of water to it,
shake well, stable froth (foam) is formed.

4.3.2. Test for tannins

a. Ferric chloride test

A small amount of test solution treat with ferric chloride solution, blue
color appears if hydrolysable tannins are present and green color appears if
condensed tannins are present.
 b. Phenazone test

To the 5 ml of aqueous extract add 0.5 gm of sodium acid phosphate.

Then warm it and filter. To the filtrate, add 2% Phenazone solution, precipitate is
formed which is often colured.

c. Gelatin test

To the test solution add 1% gelatin solution containing 10% sodium

chloride. Precipitate is formed.

4.3.3. Test for amino acids

a. Millon’s test

To the test solution, add 2 ml of millons reagent, white precipitate

indicates presence of amino acid.

b. Ninhydrin test

To the test solution, add ninhydrin solution, boil, violet color indicates
presence of amino acid.

4.3.4. Test for proteins

a. Warming test

The test solution take in a test tube and heat in boiling water bath, proteins
get coagulated.

b. Test with trichloro acetic acid

To the test solution add trichloro acetic acid, precipitate is formed.

c. Biuret test

To the test solution (2 ml) add Biuret reagent (2 ml), violet color indicates
presence of proteins.
 d. Hydrolysis test

Hydrolyze the test solution with hydrochloric acid or sulphuric acid. Then
carry out the ninhydrin test for amino acid.

e. Xanthoproteic test

To the 5ml of test solution, add 1ml of concentrated nitric acid and boil,
yellow precipitate is formed. After cooling it, add 40 %sodium hydroxide solution,
orange color is formed.

4.3.5. Glycosides

a. Keller kiliani test

The test consists of boiling about 1 gm finely powered sample with 10 ml

70% alcohol for 2 to 3 minutes. The extract is filtered. To the filtrate is added, 5 ml
water and 0.5 ml strong solution of lead acetate. Shake well and separate the filtrate.
The clear filtrate is treated with equal volume of chloroform and evaporated to yield
the extractive. The extractive is dissolved in glacial acetic acid and after cooling,
2 drops ferric chloride solution is added to it. These contents are transferred to a test
tube containing 2 ml concentrated sulphuric acid. A reddish brown layer acquiring
bluish-green color after standing is observed.

b. Legal test

The extract is dissolved in pyridine, sodium nitroprusside solution is

added to it and made alkaline-pink or red color is produced.

c. Baljet test

To the section of sample, sodium picrate solution is added. It shows

yellow to orange color.
 4.3.6. Test for cardiac glycosides

a. Keddes test

Extract the drug with chloroform, evaporate to dryness. Add one drop of
90% alcohol and 2 drops of 2% 3, 5-Di nitro benzoic acid in 90% alcohol. Make
alkaline with 20% sodium hydroxide solution, purple color is produced. The color
reaction with 3, 5-Di nitro benzoic acid depends on the presence of alpha, beta
unsaturated lactones in the aglycone.

b. Keller-killiani test (Test for deoxy sugars)

Extract the drug with chloroform and evaporate it to dryness. Add 0.4 ml
of glacial acetic acid containing trace amount of ferric chloride. Transfer to a small
test tube; add carefully 0.5 ml of concentrated sulphuric acid by the side of the test
tube. Acetic acid layer shows blue color.

c. Raymonds test

Treat the test solution with hot methanolic alkali, violet color is produced.

d. Legals test

Treat the test solution with pyridine and alkaline sodium nitroprusside
solution, blood red color appears.

e. Baljet test

Treat the test solution with picric acid or sodium picrate, orange color is
formed.

4.3.7. Test for alkaloids

The qualitative chemical tests used for detection of alkaloids are

dependent on the characters of alkaloids to give precipitates as salts of organic acids
or with compounds of heavy metals, like mercury, gold, platinum, etc. The different
reagents used are Mayer‟s reagent (potassium mercuric iodide solution) giving
cream colored precipitate. Dragendorff‟s reagent (potassium bismuth iodide
solution) giving reddish brown precipitate. Wagner‟s reagent (iodine-potassium
iodide solution) yielding reddish brown precipitate. Some alkaloids also give yellow
coloured precipitates with picric acid called as Hagner‟s reagent and picrolonic acid.
Individual alkaloid gives color or precipitate with certain specific reagent.

a. Dragendorff’s test

To 2-3 ml filtrate, add few drops of Dragendorff‟s reagent. Orange brown

precipitate is formed.

b. Mayer’s test

2-3 ml filtrate with few drops of Mayer‟s reagent gives precipitate.

c. Hager’s test

2-3 ml filtrate, add few drops of Hager‟s reagent gives yellow precipitate.

d. Wagner’s test

2-3 ml filtrate with few drops of Wagner‟s reagent gives reddish brown
precipitate.

4.3.8. Test for carbohydrates

a. Molisch’s test

The test is positive with soluble, as well as, insoluble carbohydrates. It

consists of treating the compounds with alpha napthol and concentrated sulphuric
acid which gives purple color ring at the junctions of two layer.
 b. Reduction of Fehling’s solution

To the solution of sample, equal quantity of Fehling‟s solutions A and B

is added. After heating, brick red precipitate was obtained.

4.3.9. Test for flavonoids

a. Shinoda test

To dry powder or extract, add 5 ml 95% ethanol, few drops concentrated

HCl and 0.5 gm magnesium turnings. Pink color was observed.

To small quantity of extract, add lead acetate solution. Yellow colored

precipitate was formed.

4.4. Acute toxicity study as per OECD guideline 425

In the assessment and evaluation of the toxic characters of the substance,

determination of acute oral toxicity is usually an initial step. It provides information
of health hazards likely to arise from a short-term exposure by the oral route. Acute
oral toxicity is the adverse effects occurring within a short time of oral
administration of a single dose of a substance or multiple doses given within 24h.
Data from an acute study may serve as a basis for classification and labeling. LD
(medium lethal 50 doses), oral, is a statistically derived single dose of a substance
that can be expected to cause death in 50% of animals when administered by the oral
route. The LD50 value expressed in terms of test substance per unit weight of test
animal (mg/kg). It is initial step in establishing a dosage regimen in sub chronic and
other studies and may provide initial information on the mode of toxic action of a
substance.

The concept of the up and down (UDP, stair case method) was first
designed by Dixon and Mood. In this method animals of a single sex, usually
females, with the first animal receiving a dose just below the best estimate of the
LD50. Depending on the outcome for the previous animal, the dose for the next is
increased or decreased, usually by the factor of 3.2. This sequence continues until
there is a reversal of the initial outcome (i.e., the point where an increasing dose
results in death rather than survival or decreasing dose result in survival rather than
death) then, additional animals are dosed following the up-down principle until a
stopping criterion is met. If there is no reversal before reaching the selected upper
(2000 or 5000 mg/kg) limit dose, then a specific number of animals are dosed at the
limit dose. The option to use an upper limit dose of 5000 mg/kg should be taken
only when justified by a specific regulatory need.

Healthy Wistar rats weighing between 180-220 g were used to carry out
acute oral toxicity studies by the „staircase‟ method. All successive extracts of Anjir
leaves in 0.5% tween 80 was administered orally by gavages in graduated dose to
several groups of experimental animals, one dose being used per group.
Subsequently, observations of effects were made at 0,1,2,4 and 24 h for any
mortality [48]. Ethical clearance for handling the animals is obtained from the
Institutional animal ethical committee prior to the beginning of the project work
from Institutional Animal Ethics Committee (769/2010/CPCSEA) approved the
study protocol.

4.5. Experimental Designs

4.5.1. Anti-diabetic and Anti- oxidant activity of Anjir leaves

(Alloxan-induced diabetic model)

Alloxan monohydrate was first weighed individually for each animal

according to their weight and then solubilized with 0.2 ml saline just prior to
injection. Diabetes was induced by injecting it at a dose of 150 mg/kg body weight.
intraperitonially. After 1 h of alloxan administration, the animals were given feed ad
libitum, and 5% dextrose solution was also given in a feeding bottle for a day to
overcome the early hypoglycemic phase. The animals were kept under observation
and after 48 h blood glucose was measured by glucometer. The diabetic rats
(glucose level >300 mg/dl) were separated and divided into five different groups for
experimental study, with each group containing six animals [49]
 Group I Normal Control (Saline)
Group II Diabetic control (Alloxan induced)
Group III Diabetic (Alloxan induced) + Glibenclamide (5 mg/kg)
Group IV Diabetic(Alloxan induced) + Anjir leaf Extract 200
mg/kg
Group V Diabetic (Alloxan induced) + Anjir leaf Extract 400
mg/kg

For all the animals the blood glucose level is measured on Day 0, 5, 10
and 15. The results of group IV and group V (Anjir leaf extract treated groups) is
compared with group I, II (control groups) and group III (standard drug treated
group).

Blood samples collected from all the animals are further subjected to tests
for determining Lipid profile, Hepatic glycogen level and tissue concentration of
LPO, SOD and CAT. The results of the Anjir leaf extract treated groups are
compared that with the control and standard drug treated groups.

4.6. Statistical analysis

Statistical analysis of the results was done using the statistical functions of
the Graphpad Prism 5.0 software. The results were expressed in terms of mean ±
SD. The significance of difference between mean values for the various treatments
were tested using one way analysis of variance test (ANOVA test) followed by
Dunnett Multiple Comparisons Test and the p values less than 0.05 were considered
significant [50].
 5. RESULTS AND DISCUSSION

5.1. Collection and authentication of Anjir leaves

Leaves of Anjir Leaves was collected and authenticated. The collected

leaves were dried in shade and made into coarse powder. The coarse powder of
Anjir Leaves was used for further process.

5.2. Preparation of ethanolic extracts of Anjir leaves

The methanolic extract of AnjirLeaves was prepared according to the

procedure discussed in materials and methods. The concentrated product was used in
phytochemical and pharmacological screening.

5.3. Phytochemical investigation on Anjir leaves

Thepreliminary phytochemical screening like Saponins, Tannins, Amino

acids, Proteins, Glycosides, Cardiac glycosides, Alkaloids, Carbohydrates and
Flavonoids was done with the methanolic extract of Anjir Leaves according to the
procedure. In the above chemical test the methanolic extract of Anjir Leaves showed
positive results for Saponins, Tannins, Amino acids, Proteins, Cardiac glycosides,
Alkaloids, Carbohydrates and Flavonoids except glycosides. The results of
preliminary test of methanolic extract of Anjir Leaveswere shown in Table 1.

Table 1: Phytochemical screening results of Anjir leaves

S.
PHYTOCONSTUTIENT RESULT
NO
1. SAPONINS +
2. TANNINS +
3. AMINO ACIDS +
4. PROTEINS +
5. GLYCOSIDES -
6. CARDIAC GLYCOSIDES +
7. ALKALOIDS +
8. CARBOHYDRATES +
9. FLAVONOIDS +
PRESENT = (+), ABSENT = (-)
 5.4. Acute toxicity study as per OECD guideline 425

Acute toxicity test at 3000 mg/kg of leaf extracts of Anjir Leaves

produced no mortality after 24 hours of observation. The median lethal dosage
(LD50) of the ethanolic leaves extract was greater than 3000 mg/kg body weight. The
extract did not produce any grossly negative behavioral changes such as excitement,
restlessness, respiratory distress, convulsions or coma. However, a reduction in body
weights of rats was observed. The reduction in body weight may be due to reduced
fluid and water intake, which may be secondary to feeling of fullness and loss of
appetite after administration of the extract. Despite the above side effects, the very
high value of the LD50 indicated that the extract of Anjir leaves is practically
non-toxic.

5.5 Anti-diabetic and Anti- oxidant activity of Anjir leaves

(Alloxan-induced diabetic model)

Phytochemical screening of all the extract of Anjir leavesshowed the

presence of various chemical constituents, mainly tannins, saponins and flavonoids
which may be responsible for its antidiabetic and anti-oxidant properties was shown
in Table 1. The results obtained were comparable and satisfied the standard
literature. To ascertain a scientific base for the usefulness of this plant in the
treatment of diabetes, it was decided to evaluate experimental design of antidiabetic
activity by alloxan-induced model. As expected, in the diabetic control, there was
severe hyperglycemia when compared with the normal animals. When compared
with the Diabetic control, the ethanolic extract of Anjir leaves as shown inTable 2
and Figure 5, lowered the elevated blood glucose levels. It was observed that the
standard drug Glibenclamide lowered the blood glucose level significantly bringing
it nearly back to normal, whereas ethanolic extract of Anjir leaves significantly
decreased blood serum glucose in the diabetic rats on fifth, tenth, fifteenth and
twentieths days compared with the diabetic control rat‟s blood serum glucose levels.
 In the present study, diabetic rats had lower body weights, high blood
glucose level as compared to the normal rats, which confirmed the induction of
diabetic by alloxan. In spite of the increased food consumption, loss of body weight
due to defect in glucose metabolism and excessive breakdown of tissue protein is a
characteristic condition in diabetics. The treatment with ethanolic extract of Anjir
leaves improved the average body weights of rats which indicate control over
polyphagia and muscle wasting resulted due to hyperglycemic condition.

Alloxan causes massive reduction in insulin release, through the

destruction of β-cells of the Islets of Langerhans. In our study, we have observed a
significant increase in the plasma insulin level when alloxan diabetic rats were
treated with Anjir leaves. This could be due to potentiation of the insulin effect of
plasma by increasing the pancreatic secretion of insulin from existing β-cells of
islets of Langerhans or its release from bound insulin. The significant and consistent
antidiabetic effect of Anjir leaves in alloxan diabetic rats may also be due to
enhanced glucose utilization by peripheral tissues.

Table 2: Anti Diabetic Activity of Ethanolic Extract of Anjir leaves on Alloxan

Induced Diabetic Rats

G Treatmen Day 0 Day 5 Day 10 Day 15 Day 20

roup t group
No.
I Normal 98.67±3. 92.56±6. 85.87±8. 91.23±5. 87.48±6.
control 45 23 53 89 74
II Diabetic 375.39±1 398.45±1 407.51±1 436.67±1 452.74±1
control 3.67 6.72 3.34 5.49 8.39
III Diabetic + 368.73±1 304.92±1 250.79±1 233.44±1 208.25±1
Glibenclamide 0.45 4.39 4.27 1.25 1.59
IV Diabetic + 372.21±1 364.32±0 336.12±0 343.46±1 295.54±0
Extract 200 2.34 6.53 4.98 2.22 6.74
mg/kg
V Diabetic + 377.63±1 352.38±1 316.37±1 284.79±1 253.70±1
Extract 400 2.53 5.64 6.83 0.64 2.92
mg/kg
 500

450
G
L 400
U
350
C
O300 DAY 0
S
DAY 5
E 250
DAY 10
L 200 DAY 15
E 150
DAY 20
V
E 100
L
50

0
Normal Diabetic Diabetic+ glib 5mg Diabetic+Ext 200mg Diabetic+Ext 400mg

Figure 6: Anti Diabetic Activity of Ethanolic Extract of Anjir leaves

The serum lipid profile in Table 5 and Figure 8 treated with Anjir leaves
extract returned to values nearing that of the control group. This showed that
treatment with Anjir leaves significantly improved the lipid profile in diabetic
animals and reduces the glycogen level in liver shown in Table 4 and Figure 7.
Table 3and Figure 6 show the concentration lipid peroxidation and hydroperoxides
in the liver of both control and experimental groups of rats. There was a significant
elevation in tissue lipid peroxidation and hydroperoxides in diabetic rats.
Administration of Anjir leaves to diabetic rats decreased the levels of tissue lipid
peroxidation and hydroperoxides to normal levels. The concentration of tissues
LPO, SOD and CATwere significantly decreased in diabetic rats when compared to
the control group. Administrations of Anjir leaves extract to diabetic rats tend to
bring the activities of these enzymes to near normal level.
Table 3: Antioxidant Activity of Ethanolic Extract of Anjir leaves on Alloxan
Induced Diabetic Rats

LPO SOD CAT

Gr No. of moles Amount of Moles of H2O2
Treatment
oup MDA protein required for decomposed/minute/mg
group
No. formed/mg of 50% inhibition of protein
protein

I Normal Control 2.86 7.27 93.64

II Diabetic 4.23 3.89 49.37

(Alloxan induced)
control

III Diabetic 3.12 6.53 81.35

(Alloxan induced)+
Glibenclamide

IV Diabetic 4.16 4.20 55.26

(Alloxan induced)
+ Extract 200
mg/kg

V Diabetic 3.59 5.66 72.89

(Alloxan induced)
+ Extract 400
mg/kg

100
90
80
70
60
LPO
50
40 SOD
30 CAT
20
10
0
Normal Diabetic Diabetic+ glib Diabetic+Ext Diabetic+Ext
5mg 200mg 400mg

Figure 7:Antioxidant Activity of Ethanolic Extract of Anjir leaves

Table 4: Hepatic Glycogen Level of Ethanolic Extract of Anjir leaves on
Alloxan Induced Diabetic Rats

Group Treatment group Liver

No. glycogen
(mg/g)
I Normal Control 42

II Diabetic (Alloxan induced) control 15

III Diabetic (Alloxan induced) + 36

glibenclamide

IV Diabetic (Alloxan induced) + 17

Extract 200 mg/kg

V Diabetic (Alloxan induced) + 23

Extract 400 mg/kg

45
40
35
30
25
20
15
Liver Glycogen
10
5
0

Figure 8:Hepatic Glycogen Level of Ethanolic Extract of Anjir leaves

Table 5: Serum Cholesterol Level of Ethanolic Extract of Anjir leaves on
Alloxan Induced Diabetic Rats

Group Treatment group Serum

No. cholesterol (mg/dl)

I Normal Control 97

II Diabetic (Alloxan induced) control 225

III Diabetic (Alloxan induced) + 126

glibenclamide

IV Diabetic (Alloxan induced) + Extract 194

200 mg/kg

V Diabetic (Alloxan induced) + Extract 152

400 mg/kg

250

200

150

Serum Cholesterol
100

50

0
Normal Diabetic Diabetic+ Diabetic+ExtDiabetic+Ext
glib 5mg 200mg 400mg

Figure 9:Serum Cholesterol Level of Ethanolic Extract of Anjir leaves

 5.6. DISCUSSION

The currently-available drug regimens for management of diabetes

mellitus have certain drawbacks and therefore, there is a need for safer and more
effective antidiabetic drugs. This study was undertaken to assess the antidiabetic
effect of Anjir leaves. In the present study, the oral treatment of Anjir leaves extract
decreased the blood glucose levels in diabetic rats. It has been reported that using
medicinal plant extract to treat alloxan-induced diabetic rats results in activation of
ß-cells and insulinogenic effects. Anjir leaves may also have brought about
hypoglycaemic action through stimulation of surviving ß-cells of islets of
Langerhans to release more insulin. This was clearly evidenced by the increased
levels of plasma insulin in diabetic rats treated with Anjir leaves. Since the
percentage fall in plasma glucose levels was different in models with varying
intensity of hyperglycaemia, it implies that the anti hyperglycaemic effect of that
plant is dependent on the dosage of diabetogenic agent, which in turn leads to ß-cell
destruction. A number of other plants have also been observed to exert
hypoglycaemic activity through insulin release stimulatory effects. The
concentrations of lipids, such as serum cholesterol were significantly higher in
diabetic rats than in the control group. A variety of derangements in metabolic and
regulatory mechanisms, due to insulin deficiency, are responsible for the observed
accumulation of lipids. The impairment of insulin secretion results in enhanced
metabolism of lipids from the adipose tissue to the plasma. Further, it has been
reported that diabetic rats treated with Glibenclamide shows normalised lipid levels.
Thus, the results indicate that an Anjir leaf shows insulin-like action by virtue of its
lipid lowering levels. Oxidative stress has been shown to play a role in the causation
of diabetes mellitus. Antioxidants have been shown to have a role in the alleviation
of diabetes mellitus. In diabetes mellitus, oxygen free radicals (OFRs)are generated
by stimulating H2O2. In our study, concentrations of lipid peroxides and
hydroperoxides were increased in liver of diabetic rats, indicating an increase in the
generation of free radicals. Increased lipid peroxidation in diabetes mellitus can be
due to increased oxidative stress in the cell as a result of depletion of antioxidant
scavenger systems. The present finding indicates significantly increased lipid
peroxidation of rats exposed to alloxan and its attenuation by Anjir leaves treatment.
This suggests that the protective role of Anjir leaves extracts could be due to the
antioxidative effect of flavonoids present in the leaf, which in turn act as strong
superoxide radicals and singlet oxygen quenchers. Numerous studies have revealed
lowered antioxidant and enhanced peroxidative status in diabetes mellitus. In the
current study, the LPO, SOD and CAT activities were significantly reduced in the
liverof diabetic rats. These observations emphasize the critical importance of
maintaining the antioxidant potential of the pancreatic ß-cell in order to ensure both
its survival and insulin secretion capacity during times of increased oxidative stress.
Thedecreased activities of SOD and CAT in theliverduring diabetes mellitus may be
due to the production of reactive oxygen free-radical that can themselves reduce the
activity of these enzymes.
 6. SUMMARY AND CONCLUSION

1. Preliminary phytochemical analysis report data.

2. In this experimental model, the ethanolic extract Anjir leaves (200,

400 mg/kg) with reference standard Glibenclamide 5mg/kg
significantly effective in abnormalities of enzyme profile in
experimental rats.

3. The data that Anjir leaves extract is beneficial in controlling the

blood glucose level, improves the lipid metabolism and prevents
diabetic complications from lipid peroxidation and antioxidant
systems in experimental diabetic rats.

4. This could be useful for prevention or early treatment of diabetic

disorders.

5. We conclude that the Anjir leaves have potent anti-diabetic and anti-
oxidant effects in alloxan induced diabetic rats.

6. The present investigation has also opened avenues for further

research especially with reference to the development of potent
formulation for diabetes mellitus from Anjir leaves.

7. Activity guided fractionation, formulation, and its evaluation is in

progress and will be available in short period of time.

8. Data on the short and long term adverse effects of Anjir leaves
ingestion needs to be collected.
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