Effects of Aqueous Extract of Punica Granatum Seed on Triton-X100 Induced Hypercholesterolemia in Rats

TABLE OF CONTENTS

Title page                                                                                                        i                        Approval page                                                                                                ii

Dedication                                                                                                      iii

Acknowledgement                                                                                          iv        

Table of Content                                                                                             v

List of Table                                                                                                   ix

List of Figure                                                                                                  x         

Abstract                                                                                                          xi

CHAPTER ONE: INTRODUCTION                                                                   

1.1  Background of the study                                                                         1

1.2  Justification of the study                                                                         3

1.3  Aim and Objectives                                                                                3

CHAPTER TWO: LITERATURE REVIEW       

2.1       Punicagranatum fruits                                                                       5         

2.1.2.   Botanical description of Punicagranatum seed                                 10

2.1.3.   Health benefits of Punicagranatum seed                                            10

2.2       Cholesterol                                                                                         12

2.2.1.   Hypercholesterolemia                                                                        15

2.2.2.   Description of Hypercholesterolemia                                                15

2.2.3.   Signs and symptons of Hypercholesterolemia                                   19

2.2.4.   Causes of Hypercholesterolemia                                                       20       

2.2.5.   Biomakers of hypercholesterolemia                                                  22       

CHAPTER THREE: MATERIALS AND METHODS   

3.1.      Materials                                                                                             26

3.1.1.   Punicagranatum fruits                                                                       26       

3.1.2.   Experimental animals                                                                                     26

3.1.3.   Reagent and assay kits                                                                        26       

3.2.      Methods                                                                                              27       

3.2.1    Preparation of aqueous extract of Punicagranatum seed                   27

3.2.2.   Phytochemical analysis                                                                      27       

3.2.3.   Induction of Hypercholesterolemia                                                   31

3.2.4.   Experimental Design                                                                          31

3.2.5.   Preparation of Serum                                                                          32

3.3.      Biochemical assay                                                                              33

3.3.1.   Determination of Serum Total cholesterol                                         33

3.3.2    Determination of Serum High Density Lipoprotein                          35

3.3.3.   Determination of Serum Triglycerides                                               38

3.3.4.   Determination of   Low Density Lipoprotein                                     41       

3.3.5.   Determination of Very Low Density Lipoprotein                              42

3.3.6.   Determination of Artherogenic Index                                                42       

3.3.7.   Determination of Coronary Artery Risk                                             43

CHAPTER FOUR: RESULTS                                                       

4.1.      Estimation of Total phenol content of aqueous extract of

Punicagranatum seed.                                                                                    44

4.2.      Estimation of radical scavenging 2,2diphenyl – 1-  picrylhydrazyl

of aqueous  extract of Punicagranatum seed.                                    44

4.3.      Estimation of inhibitory activity using Lipase of aqueous extract of Punicagranatum seed                                                                         45

4.4       Total Cholesterol and Triglyceride cholesterol                                            

4.5.      High Density Lipoprotein and Very low density lipoprotein           

4.6.      Low density Lipoprotein                                                                               

4.7.      Atherogenic index and Coronary risk index                                                             

CHAPTER FIVE DISCUSSION                           

Conclusion                                                                                                      57

References                                                                                                      58

Appendix                                                                                                        68

ABSTRACT

The present study was carried out to investigate the effects of oral administration of aqueous extract of Punicagranatum seed on Triton X-100 induced hypercholesterolemia in rats. Hypercholesterolemia was induced by single intraperitoneal injection of Triton X-100 and the rats were later treated with aqueous extract of Punicagranatum seed at different dosage levels (50,100 and 200mg/kg body weight) and 10ml/kg/day of standard drug (Astrovastatin) for seven days. Another group received 0.9% Normal saline (1 ml/kg/day). Animals were euthanize by anaesthetization at the end of the experiment to harvest serum used for the biochemical analysis. Aqueous extract of Punicagranatum seed extract prevented triton- induced increase in serum cholesterol, triglyceride (TG), low density lipoprotein (LDL) and decrease in high density lipoprotein (HDL) in a statistically significant manner (p < 0.05) and reversed the elevated serum LDL/HDL ratio comparably to Astrovastatin, The results demonstrated the amelioration of triton- induced hypercholesterolemia in rats by aqueous extract of Punicagranatum seed. Consequently, we can hypothesize that the part of Punicagranatum seed mediated therapeutic effects is associated with its antiatherogenic/hypolidemic component.