In Vitro Pharmacological Investigations of Aqueous Fraction of Coccinia Cordifolia Leaves

IN V

A Disser

VITRO PH

FRA

rtation subm

fulfillmen

HARMACO

ACTION O

mitted to th

nt of the req

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OLOGICA

OF COCCI

he Departme

quirements f

Sub

Samiya K

ID: 20

Departme

East W

AL INVES

CINIA COR

ent of Pharm

for the degr

bmitted B

Khondaker

010-3-70-0

ent Of Pha

West Univer

STIGATI

RDIFOLIA

macy, East

ree of Bache

By

r Rinta

048

armacy

rsity

IONS OF A

IALEAVE

West Unive

elor of Phar

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ersity, in pa

rmacy.

i

US

artial

Dedication This Research paper is dedicated to

My beloved parents,

Who are my biggest Inspirations

ii

DECLARATION BY THE CANDIDATE

I, Samiya Khondaker Rinta, hereby declare that this dissertation, entitled In vitro

pharmacological investigations of aqueous fraction of Coccinia cordifolia leaves submitted

to the Department of Pharmacy, East West University, in the partial fulfillment of the

requirement for the degree of Bachelor of Pharmacy (Honors) is a genuine & authentic research

work carried out by me under the guidance of Nigar Sultana Tithi, Lecturer and Mahbubul

Hoque Shihan, Senior Lecturer, Department of Pharmacy, East West University, Dhaka. The

contents of this dissertation, in full or in parts, have not been submitted to any other Institute or

University for the award of any Degree or Diploma of Fellowship.

----------------------------------

Samiya Khondaker Rinta

ID: 2010-3-70-048

Department of Pharmacy

East West University

Aftabnagar, Dhaka

iii

CERTIFICATION BY THE SUPERVISOR

This is to certify that the dissertion, entitled In vitro pharmacological investigations of

aqueous fraction of Coccinia cordifolia leaves is a bonafide research work done, under our

guidance and supervision by Samiya Khondaker Rinta (ID: 2010-3-70-048), in partial fulfillment

of the requirement for the degree of Bachelor of Pharmacy.

------------------------------------- -------------------------------------

Nigar Sultana TithiMahbubul Hoque Shihan

Lecturer & SupervisorSenior Lecturer & Co-supervisor Department of Pharmacy Department of Pharmacy

East West UniversityEast West University

Aftabnagar, Dhaka Aftabnagar, Dhaka

iv

ENDORSEMENT BY THE CHAIRPERSON

This is to certify that the dissertation, entitled In vitro pharmacological investigations of

aqueous fraction of Coccinia cordifolia leaves is a bonafide research work done by Samiya

Khondaker Rinta (ID: 2010-3-70-048), in partial fulfillment of the requirements for the degree of

Bachelor of Pharmacy.

---------------------------------------

Dr. Chowdhury Faiz Hossain

Professor & Chairperson

Department of Pharmacy

East West University

Aftabnagar, Dhaka

v

ACKNOWLEDGEMENTS

All praise is for Almighty for all the bounties granted to me and only with His guidance and help

this achievement has become possible.

It is my pleasure and proud privilege to express my heartiest regards and gratitude to my

respected teacher and supervisor Nigar Sultana Tithi, Lecturer, Department of Pharmacy, East

West University, for her expert supervision, constructive criticism, valuable advice, optimistic

counseling, constant support and continuous backup and encouragement throughout every phase

of the project as well as to prepare this dissertation.

I am thankful to my honorable teacher and co-supervisor, Mahbubul Hoque Shihan, Senior

Lecturer, Department of Pharmacy, East West University, for his amiability to provide me with

untiring guidance, whole hearted cooperation and for his extensive knowledge in research that

helped me in all the spheres to perform the research work.

My special thanks to Nishat Nasrin, Senior Lecturer, Department of Pharmacy, East West

University and for giving me her valuable time, constant advising, encouragement and for her kind words during my troubling moments.

I would also like to put forward my most sincere regards and profound gratitude to Dr.

Chowdhury Faiz Hossain, Professor and Chairperson, Department of Pharmacy, East West

University, for giving me the opportunity to conduct such an interesting project and for facilitating a smooth conduction of my study

I would also like to extend my thanks to all the research students in the lab, lab officers and other

staffs of the Department of Pharmacy for their help and assistance, friendly behavior and earnest

co-operation which enabled me to work in a very congenial and comfortable ambience.

I owe special thanks to my fellow research group members for their immense support and

contribution in my research work.

Last but not the least, I would like to thank my family, and friends for their care and

encouragement during my research work.

vi

CONTENTS

1. Introduction 1-21

1.1 Medicinal plants 1

1.2 Medicinal plants of Bangladesh 2

1.3 Vernacular names of Coccinia cordifolia 7

1.4 Taxonomy of C. cordifolia 7

1.5 Cucurbitaceae family 8

1.5.1 Species of Cucurbitaceae available in Bangladesh 8

1.6 Coccinia cordifolia 11

1.7 Synonyms of Coccinia cordifolia 13

1.8 Distribution of Coccinia cordifolia 13

1.9 Habitant of Coccinia cordifolia 14

1.10 Morphology of plant 15

1.10.1 Stem of C. cordifolia 15

1.10.2 Leaf of C. cordifolia 15

1.10.3 Flower of C. cordifolia 16

1.10.4 Fruit of C. cordifolia 17

1.10.5 Seed of C. cordifolia 17

1.10.6 Root of C. cordifolia 18

vii 1.11 Reproduction and dispersal of Coccinia cordifolia 18

1.12 Vegetative propagation of Coccinia cordifolia 18

1.13 Nutritional value of Coccinia cordifolia 19

1.14 Local uses of C. cordifolia in Bangladesh 19

1.15 Toxicology of C. cordifolia 21

2. Literature Review 22-38

2.1 Phytochemical constituents 22

2.2 Pharmacological properties 22

2.2.1 Anti-diabetic and hypoglycemic activity 22

2.2.1.1 In vivo study on human model 22

2.2.1.2 In vivo study on animal model 24

2.2.2 Hypolipidemic and antidyslipidemic activity 25

2.2.3 Anti-inflammatory activity 27

2.2.4 Analgesic and antipyretic activity 28

2.2.5 Anti-bacterial and anti-fungal activity 28

2.2.6 Anthelmintic activity 30

2.2.7 Cytotoxic, antitumer and pesticidal activity 30

2.2.8 Antioxidant activity 31

2.2.9 Chemoprotective activity 32

2.2.10 Antiulcer and cytoprotective activity 32

viii 2.2.11 Anti-hepatotoxic activity 33

2.2.12 Fertility inducing activity 35

2.2.13 Larvicidal, repellent and egg hatching inhibition property 35

2.2.14 Antilithiatic activity 36

Rationale and objective of the work 38

3. Methods and Materials 39-57

3.1 Collection and preparation of plant material 39

3.2 Extraction of the plant material 39

3.3 Preparation of mother solution 40

3.4 Partition of mother solution 40

3.4.1 Partition with n-hexane 40

3.4.2 Partition with carbon tetrachloride 40

3.4.3 Partition with chloroform 40

3.4.4 Partition with ethyl acetate 40

3.4.5 Collection of aqueous Fraction 42

3.5 Antioxidant activity 42

3.5.1 DPPH (1,1-diphenyl-2-picrylhydrazyl) radical test 42

3.5.1.1 Principle 42

3.5.1.2 Apparatus and reagents 43

3.5.1.3 Procedure 43-44

ix

3.5.2 Total phenolic content 44



3.5.2.3 Procedure 45-46

3.5.3 Total flavonoid content 46



3.5.3.2 Procedure 47-49

3.6 Brine shrimp lethality bioassay 49

3.6.1 Principle 49

3.6.2 Apparatus and reagents 50

3.6.3 Procedure 50-52

3.6.3.1 Preparation of sea water 50

3.6.3.2 Hatching of brine shrimp 50

3.6.3.3 Preparation of test solutions 51

3.6.3.5 Counting of nauplii 52

3.7 Antimicrobial activity by disc diffusion method 52

3.7.1 Principle 52

3.7.2 Apparatus and reagents 52

3.7.2.1 Materials 52

x

3.7.2.2 Test sample of Coccinia cordifolia 53

3.7.2.3 Test organisms 53

3.7.3 Procedure 53-57

3.7.3.1 Preparation of the medium 53

3.7.3.2 Sterilization procedure 54

3.7.3.3 Preparation of test plate 55

3.7.3.4 Preparation of discs 55

3.7.3.5 Preparation of test sample 56

3.7.3.6 Application of test samples 56

3.7.3.7 Diffusion and incubation 56

3.7.3.8 Determination of antimicrobial activity by measuring

the zone of inhibition 57

4. Results and Discussion 58-71

4.1 Antioxidant test results 58

4.1.1 DPPH test results 58

4.1.1.1 Preparation of standard curve 59

4.1.1.2 Preparation of aqueous fraction curve 60

4.1.1.3 Discussion 61

4.1.2 Total phenol content test results 62


xi 4.1.2.2 Total phenolic content present in aqueous fraction 63


4.1.3 Total flavonoid content result 64


4.1.3.2 Total flavonoid content present in aqueous fraction 65


4.2 Brine shrimp lethality bio-assay result 66

4.2.1 Preparation of standard curve 67

4.2.2 Preparation of aqueous fraction curve 68

4.2.3 Discussion 69

4.3 Antibacterial test results 70

4.3.1 Zone of inhibition of standard and aqueous fraction 70

4.3.2 Discussion 71

5. Conclusion 72

5.1 Conclusion 72

6. Reference 73-79

xii

LIST OF FIGURES

Figure 1.1: Coccinia cordifolia plant 12

Figure 1.2: Coccinia cordifolia, A: Twig with flower; B: Male flower; C:

Female flower; D: L.S. female flower; E: Male flower with

corolla split open; F: T.S. Ovary 13

Figure 1.3: Regional distribution of C. cordifolia (Synonym: Coccinia

indica) and other most frequently mentioned plants 14

Figure 1.4: C. cordifolia leaves, fruits and flowers. 15

Figure 1.5: Leaves of C. cordifolia 16

Figure 1.6: Flower of C. cordifolia 16

Figure 1.7: Fruits of C. cordifolia- (a) unripe fruit; (b) ripe fruit; (c) cross-

section of unripe fruit 17

Figure 1.8: Seeds of C. cordifolia 17

Figure 1.9: Roots of C. cordifolia 18

Figure 3.1: Drying of extract using rotary evaporator 39

Figure 3.2: Schematic representation of the partitioning of methanolic

crude extract of Coccinia cardifolia leaves 41

Figure 3.3: Mechanism of free radical scavenging activity 42

Figure 3.4: Schematic diagram of DPPH test 44

Figure 3.5: Schematic diagram of flavonoid content test 49

Figure 3.6: Artemia salina 24 hours old 51

Figure 3.7: Autoclave machine 54

Figure 3.8: Laminar hood 55

Figure 3.9: Incubator 57

Figure 4.1: Regression line and R2 value of ascorbic acid 59

Figure 4.2: Regression line and R2 value of aqueous fraction of Coccinia

xiii

cordifolia 60

Figure 4.3: Comparison between IC50 values of standard and extract 61

Figure 4.4: Graphical representation of assay of phenolic content of

ascorbic acid 62

Figure 4.5: Comparison between absorbances of standard and extract over

different concentrations 63

Figure 4.6: Graphical representation of assay of flavonoid content of

ascorbic acid 65

Figure 4.7: Plot of % mortality and predicted regression line of Tamoxifen

(standard) 67

Figure 4.8: Plot of % mortality and predicted regression line of aqueous

fraction (extract) 68

Figure 4.9: Comparison between LC50 values of standard and extract 69

Figure 4.10: Comparison of antimicrobial activity between Azithromycin

and aqueous fraction 71

xiv

LIST OF TABLES

Table 1.1: Name and medicinal uses of some common plants in Bangladesh 2

Table 1.2: Showing the vernacular names of Coccinia cordifolia 7

Table 1.3: Some plants of Cucurbitaceae family available in Bangladesh 9

Table 1.4: Nutritional value of per 100gm of edible portion of C. cordifolia 19

Table 1.5: Use of C. cordifolia in different parts of Bangladesh 19

Table 3.1: Composition of 100mg Folin-Ciocalteu Reagent 44

Table 3.2: Different concentrations of ascorbic acid solution preparation 48

Table 3.3: List of micro-organisms 53

Table 4.1: % inhibition and IC50 values of ascorbic acid 59

Table 4.2: % inhibition and IC50 values of aqueous fraction of Coccinia

cordifolia 60

Table 4.3: Free radical scavenging capacity of ascorbic acid and aqueous

fraction of Coccinia cordifolia leaves 61

Table 4.4: Total phenol content of ascorbic acid 62

Table 4.5: Total phenolic content of aqueous fraction of leaves of Coccinia

cordifolia 63

Table 4.6: Total flavonoid content of ascorbic acid 64

Table 4.7: Total flavonoid content of aqueous fraction of leaves of

Coccinia cordifolia 65

Table 4.8: Results of the bioassay of Tamoxifen (standard) 67

Table 4.9: Results of the bioassay of aqueous fraction (extract) 68

Table 4.10: Cytotoxic activity of Tamoxifen and aqueous fraction of

Coccinia cordifolia leaves 69

Table 4.11: Antimicrobial activity of standard sample (Azithromycin) and

aqueous fraction 74

xv

LIST OF ABBREVIATIONS

g microgram l microliter AAE Ascorbic Acid Equivalent

ALK.P alkaline phosphatase

ALP alkaline phosphatase

ALT alkaline aminotransferase

AST aspartate aminotransferase

DMSO dimethyl sulfoxide

DPPH 1,1-diphenyl-2-picrylhydrazyl

CI Coccinia indica

CLEt Coccinia indica leaf extract

CP cyclophosphamide

FCR Folin-Ciocalteu Reagent

g gram

GSH glutathione

hr hour

IC50 The concentration of a drug which is required for 50% of inhibition of a

specific test.

LC50 The lethal concentration required to kill 50% of the sample population of

a specific test.

LPO Lipid peroxidation

L.S. Longitudinal Section

MDA malondialdehyde

mg milligram

ml milliliter

xviPI protease inhibitor

SGOT Serum glutamic oxaloacetic transaminase

SGPT Serum glutamic pyruvic transaminase

SOD Superoxide dismutase

STZ Streptozotocin

TB Total billirubin

TP Total proteins

T.S. Transverse Section

USDA United States Department of Agriculture

UV Ultraviolet

WHO World Health Organization

xvii

ABSTRACT

The study was designed for pharmacological investigation of aqueous fraction of methanol

extract of the leaves of Coccinia cordifolia (Family: Cucurbitaceae). The powdered leaves of

Coccinia cordifolia were extracted with methanol and then partitioned with n-hexane, carbon

tetrachloride, chloroform and ethyl acetate consecutively. The aqueous fraction remaining at the

end was investigated for free radical scavenging activity (DPPH Test), total flavonoid content,

total phenol content, brine shrimp lethality test and antimicrobial test. From DPPH test the IC50

values obtained were 42.82g/ml and 25.19g/ml for standard (ascorbic acid) and aqueous fraction, respectively. The fraction contained 64.38mg AAE/g of total phenolic content and

212mg AAE/g of total flavaniod content. Screening for cytotoxic properties using brine shrimp

lethality bioassay with tamoxifen (LC50 value of 13.38g/ml) as positive control showed that the fraction have considerable cytotoxic potency exhibiting LC50 value 12.39g/ml. In antimicrobial activity investigation, the aqueous fraction showed low antibacterial and antifungal activity

against the tested organisms compared to azithromycin (30g/disc) that was used as positive

control.The aqueous fraction showed strong cytotoxic activity, moderate antioxidant activity and slight antimicrobial activity. Further investigations are needed for the proper identification and

isolation of these bioactive compounds to produce safer drugs for treatment of harmful diseases.

Key words: Coccinia cordifolia, Brine shrimp lethality bio-assay, DPPH test, phenolic content,

flavonoid content,antimicrobial activity.

In vitro pharmacological investigations of aqueous fraction of Coccinia cordifolia leaves

Chapter One INTRODUCTION

In vitro pharmacological investigations of aqueous fraction of Coccinia cordifolia leaves 1

1.1 Medicinal Plants

The use of natural products with therapeutic properties is as ancient as human civilization and,

for a long time, mineral, plant and animal products were the main sources of drugs. The

development of organic chemistry resulted in a preference for synthetic products for

pharmacological treatment. Plant derived medicines are used in selfmedication in all cultures.

Only a fraction of the worlds available plants have been studied. Discovery and use of synthetic

drugs have caused side effects or adverse reactions that were not for seen in preclinical and

clinical examinations. As a result, a resurgence of interest in the study and use of medicinal

plants has been taken place during the last two decades. As a result of modern isolation

techniques and pharmacological testing procedures, new plant drugs found their way into

modern medicine as purified substances rather than in the form of galenical preparations (Reddy

et al., 2010). Compounds such as muscarine, physostigmine, cannabinoids, yohimbine, forskolin,

colchicine and phorbol esters, all obtained from plants, are important tools used in

pharmacological, physiological and biochemical studies (Williamson et al., 1996).

Plants are valuable for modern medicine in four basic ways:

1) They are used as sources of direct therapeutic agents.

2) They serve as raw materials base for elaboration of more complex semi synthetic chemical

compounds.

3) The chemical structures derived from plant sources can be used as models for new synthetic

compounds.

4) Finally plants can be used as taxonomic markers for the discovery of new compounds

(Reddy et al., 2010).

The study of traditional human uses of plants is called Ethnobotany. It is a recognised way to

discover new effective medicines for future. In ancient Greece, plants were classified and

descriptions of them were given by scholars. It aids in the identification process. Researchers

identified in 2001 that 122 compounds that are used in modern medicine, were isolated and

identified from "ethnomedical" plant sources. The current use of the active elements of the plants

is 80% similar to those of ethnomedical use (Fabricant and Farnsworth, 2001).

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lessens b

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Family:

Name: N

Distribu

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inflamma

loss treat

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Distribu

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and diges

armacological in

c Name: Ae

Rutaceae

Bel (local nam

tion: Cultiv

art used: Fru

al uses: dys

c Name: An

Annonaceae

Ata, Sharif

name)

tion: Cultiv

art used: Fru

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burning sensa

c Name: Az

Meliaceae

Neem

tion: Plante

art used: wh

al uses: an

ations, gastr

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c Name: Alo

Liliaceae

Ghritakumari

tion: Cultiv

art used: lea

al uses: pur

stive

nvestigations of

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me), Wood A

ated all over

uit

entery, diarr

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fa (local n

ated all over

uit & leaf

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d all over Ba

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c Name: La

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tion: Cultiv

art used: bar

al uses: jau

ases

c Name: Syz

Myrtaceae

am, Kalojam

rry (english

tion: Plante

art used: wh

al uses: g

nd dysentery

c Name: He

Asclepiadac

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name)

tion: Fores

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art used: roo

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good for so

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stomachi

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c Name: Tri

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ment of the

ntic, carmin

increase app

c Name: Pu

Punicaeae

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name)

tion: Plante

art used: fru

al uses: cu

ic; astringent

c Name: Ca

Caricaceae

Pepe (local n

tion: W

esh

art used: Fru

al uses: dig

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nvestigations of

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eful in drops

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d throughou

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f aqueous fracti

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cal name);

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throughout

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of the tee

armacological in

c Name: An

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Anaras (local

tion: Cultiv

art used: lea

al uses: anth

c Name: Ad

Acanthaceae

Basak (loca

tion: Cultiv

art used: roo

al uses: exp

gonorrhoea

c Name: Cin

Lauraceae

Tejpata (loc

f (english na

tion: Cultiv

ally in other

art used: lea

al uses: stim

a, dyspepsia

eth, sore thro

nvestigations of

nanas sativus

ae

l name), Pine

ated all over

af

helmintic

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e

al name), M

ated through

ot, bark and l

pectorant, an

and rheuma

nnamomum

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af

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ion of Coccinia

lish name).

h

ut (english

desh.

c properties,

sia Lignea,

Sylhet and

ed in colic,

es, diseases



1.3 Vernacular Names of Coccinia cordifolia

Table 1.2: Showing the vernacular names of Coccinia cordifolia (Ajmal and Pandey, 2006;

Kumar et al., 2013)

Bangla Telakucha, Vinbu, Kakjhinga, Kawoaluli, Kanduri, Telakochu shag (Chakma),

Muss si (Murang)

English Ivy-Gourd, Scarlet Gourd

Hindi Kundaru ki bel, Tindora, Kovaikkai

Urdu Kunduru

Sanskrit Tundika

Assam Kawabhaturi

Malayalam Tendli, Ghiloda, Kundri, Kowai, Kovai, Kovakkai

Danish Skariagenagurk

Chinese Hong Qua

Japanese Yasai, karasuuri

Spanish Pepino, cimaron

Others Kovakka, Tindla, Kundri, Bhimb, Gentleman's Toes, Kowai, Tindori, Dondakaya,

Ghiloda, Little Gourd, Thainli, Tendli, Thendli, Manoli, Thondai

1.4 Taxonomy of C. cordifolia

Domain: Eukaryota

Kingdom: Plantae

Subkingdom: Tracheobionta

Superdivision: Spermatophyta

Division: Magnoliophyta

Class: Magnoliopsida

Order: Cucurbitales

Family: Cucurbitaceae

Genus: Coccinia

Species: Coccinia cordifolia


Botanical name: Coccinia cordifolia (Ayurveda Informatics, 2011; Pekamwar et al., 2013;

USDA, 2014)

1.5 Cucurbitaceae Family

Cucurbitaceae, the gourd family of flowering plants, belongs to the order Cucurbitales and

contains 118 genera and 845 species of food and ornamental plants. It includes the gourds,

melons, squashes, and pumpkins. Most species are prostrate or climb by tendrils. They are

annual or perennial herbs native to temperate and tropical areas. No member of the family

tolerates frost or cold soil. Most species are extremely sensitive to temperatures near freezing, a

factor that limits their geographic distribution and area of cultivation. The family includes such

economically important food plants as pumpkin, cucumber, gherkin, watermelon, muskmelon,

summer squash, winter squash, chayote, cassabanana, squash, and gourd. They are distributed in

tropical and subtropical regions (Encyclopaedia Britannica, 2014).

Members of the family are fast-growing, with long-stalked leaves that alternate along the stem.

Most species have unisexual flowers, which are borne in the leaf axils and have five white or

yellow petals. At the side of the leafstalk in annual species there is a simple, often branched,

spirally coiled tendril. It is generally regarded by most botanists to be a modified shoot. There

are five sepals in each flower; male flowers have up to five anthers, often fused or joined in a

complex way, and female flowers usually have three carpels. The fruit in most species is a

fleshy, many-seeded berry with a tough rind, often attaining considerable size. The seeds are

flattened and sometimes have beautiful wings (Encyclopaedia Britannica, 2014).

1.5.1 Species of Cucurbitaceae available in Bangladesh

Cucurbitaceae plants grow well in Bangladesh. They are found in plain areas as well as in hilly

areas like Sylhet and Chittagong. According to the recent reports of Bangladesh National

Harberium, the following Cucurbitaceae plants are available in Bangladesh as shown in table

below.

In vitro pha

Tab

Species:

Local na

Plant pa

Medicin

tapeworm

Species:

Local na

Plant pa

Medicin

against

diuretic

cures pai

cough, as

Species:

Local na

Plant pa

Medicin

renoprote

induced

ulcer and

armacological in

le 1.3: Some

Cucurbita m

ame: Mistiku

arts: Seeds

al or oth

ms

Lagenaria s

ame: Lau, K

arts: Fruits

al or other

doxorubici

activity, an

in, ulcers an

sthma and ot

Benincasa h

ame: Chalku

arts: Fruits, S

al or ot

ective activ

renal dama

d antifungal a

nvestigations of

e plants of C

maxima

umra

her uses:

siceraria

Kadu, Pani La

uses: Cardi

in induced

nti hyperlip

nd fever and

ther bronchi

hispida

umra

Seeds

ther uses

vity on isch

age, manag

activity

f aqueous fracti

Cucurbitacea

Treatment

au.

oprotective

d cardiotox

pidemic act

used for pe

al disorders

: Antidiar

hemia/reperf

gement of p

ion of Coccinia

ae family ava

t for

effect

xicity,

tivity,

ectoral

rrheal,

fusion

peptic

cordifolia leave

ailable in Ba

es

angladesh (UUddin, 2014)

9

).


Species: Cucumis sativus

Local name: Khira, Shasha

Plant parts: Fruits, Seeds

Medicinal or other uses: Removing constipation

and aid indigestion, has demulcent property,

cooling, tonic, diuretic and anthelmintic

Species: Cucumis melo

Local name: Kharmuj


Medicinal or other uses: Cooling, tonic,

laxative, aphrodisiac and diuretic; cures

biliousness, insanity, liver and kidney troubles,

bronchitis, burning of the throat, chronic fever,

painful discharges and suppression of urine

Species: Luffa aegyptiaca

Local name: Dhundul

Plant parts: Fruits

Medicinal or other uses: Expectorant, tonic,

laxative and diuretic; spleen diseases, leprosy,

piles, fever and bronchitis


Species: Momordica charantia

Local name: Korola


Medicinal or other uses: Anthelmintic,

antiemetic, carminative, purgative and for the

treatment of anemia, jaundice, malaria, cholera;

and also has antidiabetic, antioxidant property

Species: Trichosanthes cucumerina

Local name: Chichinga

Plant parts: Roots, Fruits, Seeds

Medicinal or other uses: Cure for bronchitis,

headache and boils, considered as cathartic, used

as an anthelmintic and stomach disorders.

1.6 Coccinia cordifolia

Coccinia cordifolia is one kind of species of Cucurbitaceae commonly known as telakucha, ivy

gourd, scarlet gourd, tindori, tindola, or kovai kai. It is native to north-central East Africa and is

also found wild in the Indo-Malayan region. Coccinia includes 29 additional species and they are

found only in tropical Africa (Singh, 1990).

In vitro pha

Coccinia

Australia

in these p

(Muniapp

of Southe

armacological in

a cordifolia w

a, Pacific Isl

parts of the w

pan et al., 2

east Asia (Sh

nvestigations of

Fig

was introduc

lands, the Ca

world becau

009). It is co

haheen et al.

f aqueous fracti

gure 1.1: Co

ced by huma

aribbean, an

use it is capab

onsidered as

., 2009).

ion of Coccinia

occinia cord

ans mostly as

nd southern U

ble of thrivin

s a valuable

cordifolia leave

difolia plant

s a food crop

United State

ng well in w

wild vegetab

es

p to several

es. It has be

warm, humid

able by the in

countries in

come natura

d, tropical re

ndigenous p

12

n Asia

alized

gions

eople

In vitro pha

Figure

L.S. fem

1.7 Syno

Coccinia

2011; Ku

1.8 Dist

C. cordif

Pakistan,

India in w

found in

2013).

armacological in

e 1.2: Coccin

male flower;

onyms of C

a indica; Ce

umar et al., 2

ribution of

folia is dist

, India and S

wild. It is m

areas like so

nvestigations of

nia cordifoli

E: Male flo

Coccinia co

ephalandra

2013; Pekam

f Coccinia c

tributed in A

Srilanka. It i

more common

outhern Asia

f aqueous fracti

ia, A: Twig w

ower with co

rdifolia

indica; Bryo

mwar et al., 2

cordifolia

Africa, tropi

is found in c

nly seen in a

an islands an

ion of Coccinia

with flower;

rolla split op

onia cordifo

2013).

ical Asia, a

climate that i

areas like Be

nd West Indi

cordifolia leave

; B: Male flo

pen; F: T.S.

olia; Coccin

and is comm

is warm and

engal, Bihar

ies and Haw

es

ower; C: Fem

Ovary (Sam

nia grandis

monly found

d humid. It i

and Orissa.

wain islands (

male flower;

mbamurty, 20

(Shivhare e

d in Bangla

is found in w

In generally

(Pekamwar e

13

D:

005)

et al.,

adesh,

whole

y it is

et al.,

In vitro pha

This plan

Hawaiian

Western

range of

Eastern P

througho

Figure

The urba

Azadirac

(Jaam/Ka

charantia

highest n

al., 2013

1.9 Hab

Coccinia

zones. It

adapted t

armacological in

nt has been

n and Mari

Australia, th

C. cordifoli

Papua, New

out the count

1.3: Region

freq

an and rural

chta indica

alojam tree)

a (Korola tr

number of ci

).

bitant of Co

a cordifolia

grows as a v

to tropical a

nvestigations of

n spread in

iana Islands

he northern t

a also found

w Guinea an

try in fallow

nal distributi

uently ment

l distribution

(Neem tree

, Terminalia

ree) and Swi

itations in bo

occinia cord

is a warm c

vine and can

and subtropi

f aqueous fracti

America an

s of the Pa

territory and

d in Philippin

nd Northern

lands and on

on of C. cor

ioned plants

n of C. cord

e), Trigonell

a chebula (H

ietenia mah

oth urban an

difolia

climate; shor

n be trellised

cal areas. C

ion of Coccinia

nd Pacific, b

acific. Smal

d the northern

nes, China,

n Territories

n fences (Pe

rdifolia (Syn

s in Banglade

difolia is com

la foenum-g

Haritaki tree)

agoni. It ha

nd rural regi

rt-lived pere

d when culti

C. cordifolia

cordifolia leave

but it has b

l population

n coastal par

Indonesia, M

(Australia)

ekamwar et a

nonym: Cocc

esh (Ocvirk

mpared with

graecum (M

), Ficus rac

as been foun

ions compar

ennial plant

ivated in a h

is growing

es

become inva

ns are scatt

rts of Queen

Malaysia, Th

. In Bangla

al., 2013; M

cinia indica)

et al., 2013)

h other com

Methi tree), S

cemosa (Dum

nd that C. c

red to other

that thrives

home vegetab

wild throug

asive only in

tered throug

nsland. The n

hailand, Viet

adesh it is f

BG, 2008).

) and other m

)

mmon plants

Syzygium cu

mur), Momo

cardifolia ha

plants (Ocv

s in rainy, h

ble garden. I

ghout Bangla

14

n the

ghout

native

tnam,

found

most

like,

umini

rdica

as the

irk et

humid

It has

adesh

In vitro pha

and is al

sheltered

trees. Th

up electri

1.10 Mo

C. cordif

1.10.1 S

It has ste

at the no

become s

1.10.2 L

Leaves h

smell. Th

armacological in

so cultivated

d position an

hese plants, w

icity poles, o

orphology o

folia is a dioe

Stem of C. c

ems and coil

odes. Initiall

swollen and

Leaf of C. c

have bright g

hey are trian

nvestigations of

d in various

nd a sandy s

when cultiva

over nearby

of Plant

ecious, peren

Figure 1.4

cordifolia

led tendrils.

ly, younger

semi-succul

ordifolia

green upper s

gular or pen

f aqueous fracti

parts of Ba

soil. It can fo

ated in garde

trees and int

nnial and he

4: C. cordifo

When stems

stems are

lent in nature

surface and p

ntagonal in sh

ion of Coccinia

angladesh. It

form dense m

ens, will qui

to suburban

erbaceous cli

folia leaves, f

s of C. cordi

slender, gre

e (Kumar et

pale-green u

hape (Kuma

cordifolia leave

t is an outdo

mats that rea

ickly spread

bush land (P

imber (Kum

fruits and flo

ifolia touch

een, and sm

al., 2013).

underneath, w

ar et al., 2013

es

oor plant bu

adily cover

d along fence

Pekamwar et

mar et al., 201

owers.

soil, they st

mooth but as

with charact

3).

ut prefers a s

shrubs and

es and roads

t al., 2013).

13).

trike roots re

s they grow

teristic odou

15

sunny

small

sides,

eadily

they

ur and

In vitro pha

1.10.3 F

The flow

borne sin

long) tha

tips. The

white, 3

(present a

armacological in

Flower of C

wers are acti

ngly in the le

at are joined

calyx has fi

4.5cm long

as staminode

nvestigations of

Fi

C. cordifolia

inomorphic

eaf forks on

together at t

five subulate

g, deeply d

es in female

Fi

f aqueous fracti

igure 1.5: L

a

and nearly a

stalks 1-5cm

the base and

, recurved lo

divided into

flowers). Th

igure 1.6: F

ion of Coccinia

Leaves of C.

always unise

m long. The

d usually hav

obes, each 2

five ovate

he ovary is i

lower of C.

cordifolia leave

cordifolia

exual. These

ey have five

ve five sprea

25mm long

lobes. Each

inferior (Kum

cordifolia

es

e white, tub

small narrow

ading petal lo

. The corolla

h flower ha

mar et al., 20

bular, flower

w sepals (6-

obes with po

a is campanu

as three sta

013).

16

rs are

-8mm

ointed

ulate,

amens

In vitro pha

1.10.4 F

The ivy

raw fruit

fruit (2.5

several p

Figure 1

1.10.5 S

Seeds are

grey (Shi

armacological in

Fruit of C. c

gourd fruit b

is green in c

5-6cm long a

pale, flattened

1.7: Fruits of

Seed of C. c

e obovoid an

ivhare et al.,

nvestigations of

cordifolia

belongs to th

color resemb

and up to 3

d seeds (Kum

f C. cordifol

cordifolia

nd rounded

, 2011).

F

f aqueous fracti

he berry typ

bles a small

.5cm wide)

mar et al., 20

lia-(a) unripe

at the apex,

Figure 1.8: S

ion of Coccinia

pe: oval and

dark green c

turn bright

013).

e fruit; (b) ri

, slightly pap

Seeds of C. c

cordifolia leave

hairless wit

cucumber wi

scarlet red

ipe fruit; (c)

pillose, muc

cordifolia

es

th thick and

ith paler strip

as they mat

cross-sectio

ch compress

d sticky skin

pes. These f

ture and con

on of unripe

ed and yello

17

n. The

fleshy

ntains

fruit.

owish

In vitro pha

1.10.6 R

The fresh

attached

shows ci

and thoro

2011).

1.11 Rep

This spec

from Au

self-fertil

create fru

fragment

common

dormancy

quickly,

1.12 Veg

Ivy gour

Coccinia

plant is M

The vari

armacological in

Root of C. c

h root is thic

to it. Roots

ircular outlin

ough perme

production

cies reprodu

gust to Sept

le. The plan

uit. Seeds m

ts can be sp

ly occurs by

y and usual

capable of g

getative pr

rd will grow

a cordifolia n

May-June an

iety is prop

nvestigations of

cordifolia

ck, tuberous,

s are flexible

ne and is ch

eation of par

F

n and dispe

uces by seed

tember. The

nts are dioec

may be dispe

read by floo

y humans w

ly germinate

growing 4 inc

ropagation

w in slightly

needs full su

nd Septemb

pagated thro

f aqueous fracti

, long taperi

e, soft and b

haracteristic

renchyma w

Figure 1.9: R

ersal of Coc

and also ve

species is d

cious, which

ersed by bir

ods or in dis

hen we deli

e within 24

ches per day

of Coccini

y sandy loa

un and consi

ber-October.

ugh vegetat

ion of Coccinia

ing, more or

break with a

of storage ty

with vascular

Roots of C. c

ccinia cord

egetatively v

dioecious an

h means the

rds and othe

scarded gard

iberately cul

4 weeks at 2

y (Bird, 1990

ia cordifolia

am soils wit

stent moistu

The plants

tion. Plantin

cordifolia leave

r less tortuou

a fibrous fra

ype. Parench

r elements i

cordifolia

difolia

via stem frag

nd pollinated

ey need two

er animals t

den waste. L

ltivate these

20C. Ivy g

0).

a

th good dra

ure. So, the s

should be i

ng the three

es

us with a few

acture. A tra

hyma is full

is observed

gments. C. c

d by insects.

o vines of d

that eat the

Long distanc

plants. Seed

gourd takes h

ainage. For

suitable plan

irrigated dur

e-noded cutt

w fibrous ro

ansaction of

l of starch g

(Shivhare e

cordifolia flo

. The plant i

different sex

fruit, while

ce dispersal

ds do not ex

hold and sp

the best har

nting time fo

ring hot wea

tings is the

18

otlets

f root

grains

et al.,

owers

is not

xes to

stem

most

xhibit

reads

rvest,

or this

ather.

e best


method. The selected cuttings should be planted in pits of 60cm diameter dug at a spacing of 2m

by 2m. Two or three cuttings may be planted in each pit. The most important thing is that care

should be taken to keep the root zone sufficiently moist. It is observed that manure along with

70g nitrogen and 25g each of phosphorus and potash per pit would provide the best result. The

plants should be allowed to climb over pandal or trellises (Gautam et al., 2014).

1.13 Nutritional value of Coccinia cordifolia

The nutritional value per 100gm of edible portion of Coccinia cordifolia is shown in table below.

Table 1.4: Nutritional value of per 100gm of edible portion of C. cordifolia (Gautam et al.,

2014)

Components Amount

Energy 21 (KCal)

Protein 1.4 (g)

Carbohydrate 3.4 (g)

Fat 0.2 (g)

Calcium 25 (mg)

Iron 0.9 (mg)

1.14 Local uses of C. cordifolia in Bangladesh

Table 1.5: Use of C. cordifolia in different parts of Bangladesh (Rahmatullah, 2010)

Local

Name Tribe Village District

Part(s)

Used Ailment

Nichu-

bang

Marma The fruit is given for respiratory

problems and lung disorders.

Telakucha Khasia Juice and paste of the leaves to be

taken with honey for diabetes.

Telakucha Dashuria

village

Pabna Whole

plant

Diabetes, carminative, hypertension,

fever.

Telakucha Boro bon

gram

Rajshahi Leaf,

stem

For diarrhea, juice of young leaves is

(1/32kg) is taken with a little sugar


Local


Part(s)

Used Ailment

village every morning till cure.

For blood dysentery, 1/32kg of

leaves is taken and juice squeezed

out. The juice is heated in an iron pot

and mixed with 1/64kg of sugar. To

be taken every morning on an empty

stomach.

For dizziness from sunstroke, juice

from leaves mixed with a little water

is applied to forehead and top of

head.

Telakucha Rajshahi Leaf The fried leaves are taken for

diabetes. Paste of leaves is applied to

hands or head if they feel hot.

Telakucha Muslim Rajshahi Root In case of persistent bleeding

following menstruation, the roots are

cut into small pieces and taken with

molasses sherbet.

Telakucha Kodra

village

Jessore Leaf For dysentery, leaf juice is taken

with a little salt for a week.

Telakucha Bajua

danga

village

Jessore Root For leucorrhea and menstruation

problems, pills are made from a

combination of root of Coccinia

cordifolia, root bark of Abroma

augusta, root of Achyranthes aspera,

whole plant of Ipomoea paniculata

along with honey and opium. Three

tablets are to be taken in the morning

on an empty stomach.


Local


Part(s)

Used Ailment

Telakucha Natore Leaf For cough relief and anti-pyretic

effect, 1 glass of leaf juice is taken

with sugar thrice daily.

Telakucha Narshingdi Juice from the leaves is used in

diabetes.

Telakachu Garo Sherpur For diabetes, mix enough water with

leaf and stem to make 125ml of

juice. Take two times daily (morning

before meal, and night after meal).

Kawla

kochu

pata

Noakhali Leaves The leaves are washed and crushed

to make a semi-solid paste. Pills

made from the paste are dried and

given to patients for controlling

diabetes mellitus.

Note: the housewives claim that this

preparation can cure diabetes

mellitus.

1.15 Toxicology of C. cordifolia

No toxic effect was seen on human body with C. cordifolia consumption. A study was planned to

assess the acute toxicity of Coccinia indica (Synonym: Coccinia cordifolia, Coccinia grandis)

leaves. Rats were orally administrated single dose of 100, 500 and 1000mg/kg of ethanolic

extract of Coccinia indica. Mortality, signs of toxicity, body weight, food consumption and gross

findings were observed for 14 days post treatment of Coccinia indica extract. In addition, no

significant differences were noticed in the body and organ weights between the control and

treated groups. These results state that ethanol extract of Coccinia indica is toxicologically safe

by oral administration (Baghe et al., 2011).

In vitro pharmacological investigations of aqueous fraction of Coccinia cordifolia leaves

Chapter Two LITERATURE REVIEW


2.1 Phytochemical constituents

Phytochemical screening and antimicrobial activity of Coccinia cordifolia L. plant

The medicinal plant, Coccinia cordifolia L. was analyzed in a research for its chemical

composition. Chemical analysis showed that the plant is rich in nutrients, especially antioxidant

compounds such as total phenol, vitamin C and -carotene. Phytochemical screening showed that the methanolic extract contains the bioactive constituents such as tannins, saponins, phenols,

flavonoids and terpenoids (Khatun et al., 2012).

Studies on phytochemical constituents, quantification of total phenol, alkaloid content and

In-vitro anti-oxidant activity of Coccinia cordifolia

In a study extracted dried plant of Coccinia cordifolia in hexane, ethyl acetate, ethanol (70% v/v)

and methanol was collected and these extracts were checked for their phytochemical

constituents. The whole plant of Coccinia cordifolia revealed the presence of steroids,

triterpenoids, glycosides, saponins, tannins, alkaloids, saponins, phenols and carbohydrates but

extract of Coccinia cordifolia give the negative results for quinines and oils. The methanolic

extract has more phenolic content than other extracts and the ethanol (70% v/v) extract has more

alkaloid content than other extracts (Ganga et al., 2011).

2.2 Pharmacological properties

2.2.1 Anti-diabetic and hypoglycemic activity

2.2.1.1 In vivo study on human model

Blood Sugar Lowering Effect of Coccinia grandis (L.) J. Voigt: Path for a New Drug for

Diabetes Mellitus

Double-blind phase I clinical trial was conducted at the general hospital and a private hospital in

Matara in August 2009. Sixty-one healthy volunteers were given a meal for dinner containing

20g of leaves of Coccinia grandis which was mixed with a measured amount of scraped coconut

and table salt for breakfast, and other 61 were given the placebo meal for dinner which also

contained scraped coconut and salt. They maintained a 10-hour fasting period. Glucose tolerance

test was performed blindly for the two groups. Results showed that overall blood sugar levels of

the experimental group were also significantly lower than those of the control group (F(1,117)


5.56, P < 0.05). Increase in the blood sugar levels from fasting to one hour (F(1,117) 6.77, P <

0.05) and two hours (F(1,117) 5.28, P < 0.05) postprandially was statistically significant for

participants who were in the control group than those of in the experimental group. The mean

difference of postprandial blood sugar levels (mg/dL) after one hour (20.2, 95% confidence

interval, 4.81 to 35.5) and two hours (11.46, 95% confidence interval; 1.03 to 21.9) was

statistically significant between the two groups. This proved that Coccinia grandis has a blood

sugar lowering effect (Munasinghe et al., 2011).

Effect of Supplementation of Coccinia cordifolia Extract on Newly Detected Diabetic

Patients

This study aimed to evaluate the effectiveness of Coccinia cordifolia on blood glucose levels of

incident type 2 diabetic patients requiring only dietary or lifestyle modifications. A double blind,

placebo control, randomized study trial was carried out. Sixty incident type 2 diabetics (aged 35

60 years) were recruited. The subjects were randomly assigned into the placebo or experimental

group and were provided with 1g of an alcoholic extract of the herb for 90 days. Anthropometric,

biochemical, dietary and physical activity assessment were carried out at baseline and were

repeated at day 45 and day 90 of the study. All the subjects were provided with standard dietary

and physical activity advice for the control of their blood sugars. Results showed a significant

decrease in the fasting, post prandial blood glucose and glycosylated hemoglobin of the

experimental group when compared to the placebo group. The fasting and post prandial blood

glucose levels of the experimental group at day 90 significantly decreased by 16% and 18%

respectively. This study suggests that Coccinia cordifolia extract has a potential hypoglycemic

action in patients with mild diabetes (Kuriyan et al., 2008).

Coccinia indica in the treatment of patients with diabetes mellitus

A double blind control trial was conducted with preparation from the leaves of the plant,

Coccinia indica, on uncontrolled, maturity onset diabetics. The trial lasted for six weeks for an

individual patient. Out of the 16 patients who received the experimental preparations 10 showed

marked improvements in their glucose tolerance while none out of the 16 patients in the dummy

group showed such a marked improvement. This difference is highly significant (kappa 2 with

Yates' correction = 11.7, P < 0.001) (Khan et al., 1979).


2.2.1.2 In vivo study on animal model

Comparative efficacy of Telakucha (Coccinia indica) leaves and Amaryl(R) Tablet

(Glimepiride) in induced diabetes mellitus in rat

Thirty healthy rats of both sexes weighing between 150 to 200gm were selected from among the

offspring and randomly divided into six equal groups. Rats of Group A and Group B were kept

as non-hypoglycemic control and hypoglycemic control, respectively. After acclimatization

hyperglycemia was induced in five groups of rats (B, C, D, E and F) by administering

streptozotocin (STZ) intraperitoneally at a dose of 55mg/kg body weight. After fifteen days of

STZ injection, four groups of rats (C, D, E and F) were administered with Telakucha (Coccinia

indica) and Amaryl(R) (Glimepiride) as per schedule dose and all the control and treated rats were

closely observed during 14 days of treatment. The oral administration over 14 days of Telakucha

(Coccinia indica) leaves extract significantly lowered blood glucose level but was not as potent

as patent drug Amaryl(R) (Glimepiride). The herbal preparation also increased body weight but

not to the extent caused by the patent drug Amaryl(R). The Telakucha leaves at 750mg/kg body

weight significantly reduced (30.73%) the blood glucose level from 31.240.36mmol/L to

21.640.17mmol/L and significantly increased (5.45%) the body weight from 181.9621.10g to

191.8712.42g (Amanullah et al., 2008).

Effect of Ethanol Extract of Coccinia grandis Lin leaf on Glucose and Cholesterol Lowering

Activity

Glucose and cholesterol lowering effect of the ethanol extract of C. grandis leaf was evaluated

using the alloxan-induced diabetic rat and compared the activity with diabetic control and

antidiabetic drug (Glibenclamide). Ethanol extract (25mg/kg) of C. grandis and Glibenclamide

were administered to normal and experimental diabetic rats for the duration of 10 days. In the

alloxan-induced diabetic rat model, C. grandis (25mg/kg) significantly (p


Protective effect of Coccinia indica on changes in the fatty acid composition in

streptozotocin induced diabetic rats

The present study was undertaken to investigate the effect of Coccinia indica on blood glucose,

plasma insulin, cholesterol, triglycerides, free fatty acids and phospholipids and fatty acid

composition of total lipids in liver, kidney and brain of normal and streptozotocin (STZ) diabetic

rats. Oral administration of the ethanolic extract of Coccinia indica leaves (200mg/kg body

weight, CLEt) for 45 days to diabetic rats decreased the concentrations of blood glucose whereas

plasma insulin was elevated. These results suggest that CLEt exhibits hypoglycaemic effect in

STZ induced diabetic rats. The effect of CLEt at 200mg/kg body weight was better than that of

glibenclamide. The results indicate that the administration of CLEt to diabetic animals

normalizes blood glucose and causes marked improvement of altered carbohydrate metabolic

enzymes during diabetes (Pari and Venkateswaran, 2003).

2.2.2 Hypolipidemic and antidyslipidemic activity

Protective effect of Coccinia indica on changes in the fatty acid composition in

streptozotocin induced diabetic rats

The present study was undertaken to investigate the effect of Coccinia indica on cholesterol,

triglycerides, free fatty acids and phospholipids and fatty acid composition of total lipids in liver,

kidney and brain of normal and streptozotocin (STZ) diabetic rats. Oral administration of the

ethanolic extract of Coccinia indica leaves (200 mg/kg body weight, CLEt) for 45 days to

diabetic rats decreased the concentrations of lipids and fatty acids, viz., palmitic, stearic, and

oleic acid whereas linolenic and arachidonic acid were elevated. These results suggest that CLEt

exhibits hypolipidaemic effects in STZ induced diabetic rats. It also prevents the fatty acid

changes produced during diabetes. The effect of CLEt at 200 mg/kg body weight was better than

that of glibenclamide (Pari and Venkateswaran, 2003).

Antidyslipidemic activity of polyprenol from Coccinia grandis in high-fat diet-fed hamster

model

Ethanolic extract was fractionated into chloroform, n-butanol and water-soluble fractions and

were evaluated. Activity was proved to be concentrated in chloroform-soluble fraction.

Chloroform-soluble fraction containing active component was subjected to repeated column

chromatography, furnished a polyprenol characterized as C(60)-polyprenol(1) isolated for the


first time from this plant. It significantly decreased serum TG by 42%, total cholesterol (TC)

25% and glycerol (Gly) 12%, accompanied HDL-C/TC ratio 26% in high-fat diet (HFD)-fed

dyslipidemic hamsters at the dose of 50mg/kg body weight. Results are comparable to standard

drug, fenofibrate at the dose of 108mg/kg. So the compound polyprenol(1) isolated from leaves

of C. grandis possess marked antidyslipidemic activity (Singh et al., 2007).

Ameliorative potential of Coccinia grandis extract on serum and liver marker enzymes and

lipid profile in streptozotocin induced diabetic rats

The present study was undertaken to evaluate the potential of Coccinia grandis extract on serum

and liver marker enzymes (ALP, AST, ALT and LDH) and lipid profile (total cholesterol,

phospholipids, triglycerides and free fatty acids in serum and liver) in streptozotocin induced

diabetic animals. The experimental animals were treated with methanolic extract of Coccinia

grandis and the levels of marker enzymes and lipid profile were estimated. The ALP, AST, ALT

and LDH levels were increased in diabetic rats and restored to near normal levels after

administration of plant extract. The lipid profile increased in diabetic group and after the

treatment with the plant extract the levels were reverted to near normal. Thus the methanolic

extract of Coccinia grandis has a potent ability to restore the marker enzymes and the lipid

profile was reverted to near normal levels (Krishnakumari et al., 2011).

Effect of Ethanol Extract of Coccinia grandis Lin leaf on Glucose and Cholesterol Lowering

Activity

Glucose and cholesterol lowering effect of the ethanol extract of C. grandis leaf was evaluated

using the alloxan-induced diabetic rat and compared the activity with diabetic control and

antidiabetic drug (Glibenclamide). Ethanol extract (25mg/kg) of C. grandis and Glibenclamide

were administered to normal and experimental diabetic rats for the duration of 10 days. C.

grandis extract (25mg/kg) produced significant (p


2.2.3 Anti-inflammatory activity

Anti-inflammatory, analgesic and antipyretic activity of aqueous extract of fresh leaves of

Coccinia indica

This study was aimed to evaluate both post- and pre-treatment anti-inflammatory activities of the

aqueous extract of fresh leaves of Coccinia indica in rats using the carrageenan-induced paw

oedema method at various dose levels. Ceiling effect of the extract was observed at 50mg/kg in

pre-treatment carrageenan test. In post-treatment studies, a dose-dependent anti-inflammatory

effect was observed in the dose range of 25-300mg/kg. The effect was equivalent to diclofenac

(20mg/kg) at 50mg/kg but it was significantly pronounced at higher doses. Effectiveness of

extract in the early phase of inflammation suggests the inhibition of histamine and serotonin

release. In conclusion, this study has established the anti-inflammatory activity of C. indica and,

thus, justifies the ethnic uses of the plant (Niazi et al., 2009).

Evaluation of Anti-inflammatory activity of Coccinia indica leaves extracts

The effects of Coccinia indica leaves extracts on different phases of acute inflammation were

examined. Investigations were performed using different phlogistic agents-induced paw edema

viz.Carrageenan-induced paw oedema and Dextran- induced paw oedema in rats. Various

extracts (ethanol and aqueous) of Coccinia indica leaves extracts at a dose of 250mg/kg and

500mg/kg orally were tested. Diclofenac sodium at the dose of 10mg/kg was used as standard.

Both the extracts showed significant activity (*p


2.2.4 Analgesic and antipyretic activity

Anti-inflammatory, analgesic and antipyretic activity of aqueous extract of fresh leaves of

Coccinia indica

Analgesic and antipyretic properties were evaluated using tail flick model and yeast-induced

hyperpyrexia, respectively. The extract produced marked analgesic activity comparable to

morphine at 300mg/kg, which suggests the involvement of central mechanisms. A significant

reduction in hyperpyrexia in rats was also produced by all doses of extract with maximum effect

at 300mg/kg comparable to paracetamol. In conclusion, this study has established the analgesic

and antipyretic activity of C. indica (Niazi et al., 2009).

2.2.5 Anti-bacterial and anti-fungal activity

Antimicrobial activity of the fruit extracts of Coccinia indica

The bioactive compounds of fruits of Coccinia indica were investigated for antibacterial activity

against some pathogenic bacteria. The aqueous extracts did not show much significant activity,

while the organic extracts (petroleum ether and methanol) showed the highest activity against the

test bacteria. The activity was more pronounced on gram-positive organisms with

Staphylococcus aureus being more susceptible and Salmonella paratyphi A being more resistant

(Shaheen et al., 2009).

Antibacterial Activity of the Leaves of Coccinia indica (W. and A) Wof India

The aim of the present research was focused on investigating the antibacterial properties of

Coccinia indica (W.A.) via in vitro approach. The aqueous and organic solvent (Petroleum ether,

chloroform and ethanol) extracts from the leaves of Coccinia indica (Cucurbitaceae) were tested

against Enterobacter aerogenes, Pseudomonas aeruginosa, Staphylococcus epidermidis, Bacillus

subtilis and Salmonella typhimurium by agar well diffusion method and broth dilution method.

Results showed promising antibacterial activity against the bacteria tested. Among these, ethanol

and aqueous extracts were found to have a more potent inhibitory effect comparing with the

other extracts. This proves that there is potentiality in the plant extracts for the treatment of

various skin and gastrointestinal infections in humans (Hussain et al., 2010).


Antibacterial, cytotoxic and antioxidant activity of chloroform, n-hexane and ethyl acetate

extract of plant Coccinia cordifolia

Disc diffusion technique was used for in vitro antibacterial screening against gram positive and

gram negative human pathogenic bacteria. Here kanamycin disc (30g/disc) was used as

standard. The chloroform extract of Coccinia cordifolia showed good antibacterial activity with

the average zone of inhibition 9-12mm. The n-hexane and ethyl acetate extract showed average

zone of inhibition 7-10mm and 7-11mm respectively (Bulbul et al., 2011).

Antibacterial activity of Coccinia grandis leaf extract on selective bacterial strains

To assess the antibacterial activities of Coccinia grandis leaf extract on selective bacterial strains

under in-vitro conditions. The antibacterial activity was tested against five bacterial strains by

agar well diffusion method. The crude extract showed a broad spectrum of antibacterial activity

by inhibiting both the gram positive and gram negative groups. The antibacterial activity of C.

grandis leaf extract using solvents such as acetone, ethanol, methanol, aqueous and hexane was

evaluated against five bacterial sp. Ethanol leaf extract of C. grandis showed high antibacterial

activity against S. aureus, B. cereus, E coli, K pneumoniae and S. pyogens. Minimal inhibitory

concentration of the leaf extract against each test organism was also studied by observing their

growth on Mueller Hinton Agar containing the extract at various incremental levels, equivalent

to 31.25g/ml to 1000g/ml of the extract. The highest activity was observed in ethanol extracts against S. aureus, E. coli, and K. pneumoniae with an inhibitory concentration below 31.5g/ml. The significance of the study was conducted to investigate the in vitro antibacterial activity of

folklore medicinal plant and to evaluate scientific base of their applications (Sivaraj et al., 2011).

Antimicrobial activity of protease inhibitor isolated from leaves of Coccinia grandis (L.)

Voigt.

A 14.3kDa protease inhibitor PI exhibited marked growth inhibitory effects on colon cell lines in

a dose-dependent manner. PI was thermostable and showed antimicrobial activity without

hemolytic activity. PI strongly inhibited pathogenic microbial strains, including Staphylococcus

aureus, Klebsiella pneumoniae, Proteus vulgaris, Eschershia coli, Bacillus subtilis and

pathogenic fungus Candida albicans, Mucor indicus, Penicillium notatum, Aspergillus flavus

and Cryptococcus neoformans. Examination by bright field microscopy showed inhibition of

mycelial growth and sporulation. Morphologically, PI treated fungus showed a significant


shrinkage of hyphal tips. Reduced PI completely lost its activity indicating that disulfide bridge

is essential for its protease inhibitory and antifungal activity. Results reported in this study

suggested that PI may be an excellent candidate for development of novel oral or other anti-

infective agents (Satheesh and Murugan, 2011).

Phytochemical screening and antimicrobial activity of Coccinia cordifolia L. plant

The antimicrobial activities of the methanol, water, ethanol and ethyl acetate extracts of Coccinia

cordifolia L. plant were evaluated against some Gram positive bacteria (Sarcina lutea, Bacillus

subtilis and Staphylococcus aureus), Gram negative bacteria (Salmonella typhi, Shigella

dysenteriae and Escherichia coli) and fungi (Candida albicans, Aspergillus niger and

Penicillium notatum). In the methanolic extract of the plant, promising antimicrobial potential

was observed against the tested microorganism. Methanolic extract showed highest activity

against Shigella dysenteriae, Escherichia coli, Staphylococcus aureus, and Candida albicans

compared to the other extracts. Water extract showed less antimicrobial activity as compared to

other extractants (Khatun et al., 2012).

2.2.6 Anthelmintic Activity

Evaluation of Anthelmintic Activity of Coccinia indica (fruits)

The present study was designed to explore the anthelmintic activity of different extracts of plant

Coccinia indica (fruits) using petroleum ether, ethyl acetate methanol and water as solvents.

Various concentrations (25 and 50mg/ml) of all the extracts were tested, which involved

determination of time of paralysis and time of death of the worms. It was compared with

Albendazole as standard reference and normal saline as control. The study indicated the potential

usefulness of Coccinia indica against earthworm infections (Shivhare et al., 2011).

2.2.7 Cytotoxic, antitumer and pesticidal activity



The main aim of this study was to find out the antibacterial, cytotoxic and antioxidant activity of

chloroform, n-hexane and ethyl acetate extracts of Coccinia cordifolia (Family: Cucurbitaceae).

The Brine shrimp lethality bioassay method was used to determine the cytotoxic activity and

vincristine sulphate was used as positive control. The LC50 values of standard vincristine


sulphate, chloroform, n-hexane and ethyl acetate extract were 7.55g/ml, 23.96g/ml, 14.12g/ml & 15.49g/ml respectively which indicate the plants extracts compounds are promisingly cytotoxic and they might have antitumer and pesticidal activity. Among the extracts

the chloroform extract showed the highest cytotoxic activity with LC50 23.96g/ml (Bulbul et al., 2011).

2.2.8 Antioxidant activity

Effect of Coccinia indica leaf extract on plasma antioxidants in streptozotocin- induced

experimental diabetes in rats

The present study was carried out to investigate the antioxidant effect of an ethanolic extract of

Coccinia indica leaves. Oral administration of Coccinia indica leaf extract (CLEt) (200mg/kg

body weight) for 45days resulted in a significant reduction in plasma thiobarbituric acid reactive

substances, hydroperoxides, vitamin E and ceruloplasmin. The extract also caused a significant

increase in plasma vitamin C and reduced glutathione, which clearly shows the antioxidant

property of CLEt (Venkateswaran and Pari, 2003).



Antioxidant activity test of the crude extracts were assessed by means of DPPH free radical

scavenging method where ascorbic acid was used as standard with IC50 43.22 g/ml. The ethyl acetate fraction of Coccinia cordifolia showed strongest antioxidant activity with IC50 value of

50.98g/ml (Bulbul et al., 2011).

Studies on phyto chemical constituents, quantification of total phenol, alkaloid content and

In-vitro anti-oxidant activity of Coccinia cordifolia

In the present study dried plant of Coccinia cordifolia was extracted in hexane, ethyl acetate,

ethanol (70%v/v) and methanol. The selected plant extracts were produced concentration

dependent percentage inhibition of superoxide radical and produced maximum activity at a

concentration of 160g and there after the percentage inhibition were raised gradually to its maximum level with higher concentrations. Among the four types of C. cordifolia extracts, the

methanolic extract showed better activity than remaining extracts at 160g concentrations (Ganga et al., 2011).


2.2.9 Chemoprotective activity

Chemoprotective potential of Coccinia indica against cyclophosphamide-induced toxicity

The present study was aimed to investigate the chemoprotective potential of Coccinia indica

against CP-induced oxidative stress, genotoxicity, and hepatotoxicity. Rodents were orally pre-

treated with Coccinia indica extract (200, 400, and 600mg/kg) for five consecutive days. On 5th

day, these animals were injected with CP (5mg/kg i.p) and sacrificed after 24hrs for the

evaluation of oxidative stress, hepatotoxicity, micronucleus formation, and chromosomal

aberrations. It was found that the CP significantly increased malondialdehyde (MDA) and

decreased catalase and glutathione (GSH) levels in brain, and it was significantly reversed by

Coccinia indica extract (400 and 600mg/kg). Further, pre-treatment with Coccinia indica extract

(200, 400, 600mg/kg) significantly and dose-dependently reduced micronuclei formation and

incidence of aberrant cells. It was also found that the CP-induced increase in the serum

biomarker enzymes like alkaline phosphatase (ALP), alkaline aminotransferase (ALT), and

aspartate aminotransferase (AST) were significantly reduced by Coccinia indica extract. Thus,

the results indicate the protective effect of Coccinia indica extract against CP-induced oxidative

stress, genotoxicity, as well as hepatotoxicity (Nitharwal et al., 2013).

2.2.10 Antiulcer and cytoprotective activity

Antiulcerogenic and antioxidant effects of Coccinia grandis (Linn.) Voigt leaves on aspirin-

induced gastric ulcer in rats

The effect of Coccinia grandis (Linn.)Voigt leaves powder, its methanol and aqueous extracts

were investigated on aspirin-induced gastric ulcer model in rats. The leaf powder showed a

significant dose related decrease in ulcer index, with significant increase in mucus secretion and

decrease in level of Lipid peroxidation (LPO) and Superoxide dismutase (SOD) activity.

Methanol extract at an equivalent dose to that of the powder also showed a significant decrease

in ulcer index with significant changes in mucus secretion, LPO and SOD. However, aqueous

extract was found to be non-significant in reducing ulcer index. The group, receiving standard

drug Famotidine, showed no effect on the mucus secretion induced in this experimental model.

These observations confirm the antiulcerogenic potential of this plant, probably due to increased

mucus secretion and antioxidant property (Mazumder et al., 2007).


Anti-ulcer activity of ethanolic, aqueous and total aqueous extracts of Coccinia grandis

Linn. Voigt in pyloric ligature induced ulcers in albino rats

The present study was designed to investigate the antiulcer potential of ethanolic, aqueous and

total aqueous extracts of Coccinia grandis Linn. Ulcer was induced by pylorus ligature in Wistar

albino rats. Drugs were administered in two different dose levels (200mg/Kgbwt,

400mg/Kgbwt). Though all three extracts of Coccinia grandis, dose dependently reduced, the

total acidity, ulcer index, and increased pH of gastric juice, ethanol extracts exhibited markedly

significant results. However, ethanol extract has shown (78.57%) a highly significant ulcer

curative potential and decreased ulcer formation also. A preliminary phytochemical analysis

revealed the presence of different phytoconstituents such as alkaloids, carbohydrate, glycosides,

phyto sterol, saponins, volatile oil, tannins etc. which may impart their anti-ulcer activity by

acting as anti-secretory and cytoprotective agents. The present result suggests that both antisecretory and cytoprotective mechanisms of different extracts of Coccinia grandis exerted

protective effect (Santharam et al., 2013).

2.2.11 Anti-hepatotoxic activity

Antihepatotoxic activity of Coccinia indica

Aqueous, light petroleum, chloroform, alcohol, benzene and acetone extracts of the leaves of

Coccinia indica (Family: Cucurbitaceae) were screened for antihepatotoxic activity.The extracts

were given after the liver was damaged with CCl4. Liver function was assessed based on liver to

body weight ratio, pentobarbitone sleep time, serum levels of transaminase (SGPT, SGOT),

alkaline phosphatase (ALP) and bilirubin. Alcohol and light petroleum extracts were found to

have good anti-hepatotoxic activity (Gopalakrishnan et al., 2001).

Protective effect of Coccinia indica leaf extract against alcohol combined with carbon

tetrachloride and paracetamol induced liver damagein rats

The aim of our study was to investigate the effect of leaf extract of Coccinia indica against

Alcohol combines with CCl4 and Paracetamol induced hepatotoxicity. The effects of oral

treatment with Coccinia indica (CI) leaf extracts (100mg/kg and 200mg/kg for 7 days) were

studied on hepatic damage induced by alcohol (40% alcohol 2.0ml/100g, p.o. for 21 days) and

CCl4 (0.1ml/kg, s.c. on 20th day) and also with paracetamol (750mg/kg ip.) in rats. Biochemical

parameters in serum like glutamate oxaloacetate transaminase (SGOT), total billirubin (TB),


alkaline phosphatase (ALP) and total proteins (TP) were estimated to assess the liver function.

Alcohol CCl4 and paracetamol treatment produced an increase in SGOT, ALP, Total billirubin

and decrease in total proteins indicating the liver damage. These effects were progressively

reduced (SGOT, ALP and Total billirubin) and increased (Total proteins) by treatment doses of

(100mg/kg and 200mg/kg) CI leaf extracts. These biochemical observations were supplemented

by histopathological examination of liver sections. CI leaf extract protected the liver from

alcohol-CCl4 and paracetamol induced hepatic damage (Maheswari et al., 2011).

Evaluation of hepatoprotective activity of ethanol extract of Coccinia grandis (L.)

Voigt.leaves on experimental rats by acute and chronic models

The aim of this work was to study the hepatoprotective effect of crude ethanolic extract from the

leaves of C. grandis against liver damage induced by Paracetamol and CCl4 in rats.

Administration of paracetamol (750mg/kg/day) and CCl4 (3ml/kg/day) showed a marked

increase in SGOT, SGPT, ALP, bilirubin (total, direct), total proteins, globulin, cholesterol and

decrease in albumin in comparison with the normal control group. The effect of ethanol extract

of C. grandis at 150mg/kg and 300mg/kg doses reduced the serum activities caused by

paracetamol and CCl4, which were observed to be statistically significant when compared with

that of the control group. Silymarin provided a better inhibition or exhibition of the biochemical

parameters induced by paracetamol and carbon tetrachloride in rats. The activity may be due to

the presence of either alkaloids or triterpenoids or reducing sugars or their combinations, as

obtained from the preliminary phytochemical screening of the leaves of the plant. The extract

showed no signs of acute toxicity up to a dose level of 3.2gkg1 in rats by oral route. Thus, it

could be concluded that ethanolic extract of C. grandis leaves possessed significant

hepatoprotective activity (Kundu et al., 2012).

Hepatoprotective Activity of Aqueous Fruit Extract of Coccinia indica against Paracetamol

Induced Hepatotoxicity in rats

To evaluate the hepatoprotective activity of the aqueous fruit extract of Coccinia indica against

paracetamol induced hepatotoxicity in albino rats. Hepatotoxicity was induced in albino rats by

p.o of paracetamol (2gm/kg for 3 days). The aqueous fruit extract of Coccinia indica was

administered to the experimental animals at two selected doses for 14 days. The hepatoprotective

activity of the extract was evaluated by the liver function marker enzymes in the serum


(asparitate transaminases AST, alanine transaminase ALT, alkaline phosphatase ALK.P, total

bilirubin TB, and histopathological studies of liver. Both the treatment groups showed

hepatoprotective effect against paracetamol induced hepatotoxicity by significantly restoring the

levels of serum enzymes to normal which was comparable to that of silymarin group. The oral

administration of Coccinia indica significantly ameliorate

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In Vitro Pharmacological Investigations of Aqueous Fraction of Coccinia Cordifolia Leaves