Oxidative And Biochemical Parameter Assessment For Alloxan Induced Diabetic Rats Treated With Methanol Leaf Extracts Of Ocimum Gratissimum.

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ABSRACT

Ocimum gratissimum is a vegetable plant of wide nutritional and medicinal application in Nigeria and some other parts of the world. The aim of this study is to know the effect and efficacy of oral administration of methanol extract of Ocimum gratissimum on the level of oxidative and biochemical parameters of alloxan-induced diabetic rats. Leaves of Ocimum gratissimum were obtained and dried under room temperature, after drying, it was grind into powder and 320.64g soaked in 50ml of methanol for 72hours. After which it was filtered with a Whatsman (no 1) filter paper. The filtrate was concentrated using rotary evaporator at 37oC. A total of 20 male Wistar albino rats were grouped into the normal group, diabetic not treated, diabetic treated with gilbenclamide and the group treated with the leaf extract, they were all fed orally with the extract for 14days.  All the rats were sacrificed and their blood was collected for assaying of biochemical parameters. Result of the study showed a significant (P>0.05) increase in the serum SOD activity compared with the normal control.Vitamin E was observed showing a significant (P>0.05) increase in the group treated with the extracts compared with diabetic untreated. The result also showed a significant (P<0.05) decrease in MDA level of test group compared with the diabetic untreated and a non-significant (P>0.05) increase was seen in the HDL level of the test group and also a significant (P<0.05) decrease was observed in the LDL level of the group treated with leaf extract. Data of the study suggests that the oral administration of the aqueous methanol leaf extract of Ocimum gratissimum may impair naturally generated oxidant/toxicant activity and hereby enhance specific activities of antioxidants in rats.

TABLE OF CONTENTS

Title Page                                                                                                    i

Approval                                                                                                    ii

Certification                                                                                                         iii

Dedication                                                                                                  iv

 Acknowledgement                                                                                     v

Abstract                                                                                                      vi      

Table of contents                                                                                        vii

List of tables                                                                                               viii    

List of figures                                                                                             ix      

CHAPTER 1: INTRODUCTION  

1.1 Background of study                                                                                      1

1.2 Statement of the problem                                                                     2       

1.3 Aim and Objectives                                                                              2

1.3.1 Aim of the study                                                                                2

1.3.2 Objective of the study                                                                        3

CHAPTER 2: LITREATURE REVIEW

    1. About the plant                                                                                 4
      1. Scientific classification                                                                      5
      2. Chemical constituents of plant                                                                   6
      3. Geographical distribution                                                                           6
      4.  Composition of aqueous extract of Ocimum gratissimum                          6
      5.    Medical uses of Ocimum gratissimum                                            7
      6. Alternative and complimentary medical uses                                             7
      7.   Published pharmacological properties of O. gratissimum                         8
      8.  Alloxan induced diabetes                                                                 8
    2. Alloxan mechanism of action                                                          8

2.2.1 Chemical properties of Alloxan                                                                  10

2.2.2 Impact of Alloxan upon beta cells                                                     10

CHAPTER 3: METHOD AND MATERIAL

    1. Materials and methods                                                                      12
    2. chemical and reagents                                                                       12

3.1.2 Equipment                                                                                         12

3.1.3 Materials                                                                                           12     

3.2  The plant                                                                                             13

3.2.1Extract Preparation                                                                                      31

3.2.2 Preparation of extract for administration                                          14

3.2.3 Experimental animal                                                                         14

3.2.4  Experimental design                                                                         15

3.2.5 Collection of blood sample                                                                15

3.2.6 Determination of the biochemical parameter                                               15

3.2.7 Glucose level                                                                                     15

3.2.8  Determination of Malondialdehyde (MDA) level 16

3.2.9 Superoxide Dismutase (SOD) Assay                                                 18

3.3 Determination of High Density Lipoprotein (HDL) level                     20

3.3.1 Low density lipoprotein LDL                                                            21

3.3.2 Vitamin E level                                                                                  21

3.3.3 Statistical Analysis                                                                                     23

CHAPTER 4: RESULTS 

4.1  Results                                                                                                 24

4.2  Effects of Methanol extract of O.gratissimum on malondialdhyde

 (MDA)                                                                                                       25

4.3Effects of Methanol extract of O.gratissimumon superoxide dismutase

(SOD)                                                                                                         26

4.4 Effects of Methanol extract of O.gratissimumof high density

 lipoprotein (LDL) level                                                                                       27

4.5 Effects of Methanol extract of O.gratissimumof low density

lipoprotein (LDL)                                                                                       28

4.6  Effects of Methanol extract of O.gratissimumon vitamin E level                   29

CHAPTER 5 DISCUSSION AND CONCLUSION

5.1    Discussion                                                                                         30

5.2      Conclusion                                                                                                32

  References                                                                                                 33

Appendix                                                                                                   36

 Appendix

CHAPTER ONE

INTRODUCTION

    1.  BACKGROUND OF STUDY

The use of Ocimum gratissimum leaf extract to treat Alloxan induced diabetic rat and its oxidative and biochemical parameter assessments.

            In Nigeria, especially in the southern part there is a consumption of plants extracts people consume. They also consume a lot of vegetable in their native diet and some of these plants are believed to cure some aliments. Ocimum gratissimum is one of the many found to lower glycaemia in Type-1 diabetes.

            Many components of food such as secondary plants metabolites have been seen to alter biological processes which may reduce the risk of some chronic diseases in human such as diabetes.

            Diabetes mellitus has its complications especially diabetes conditions among infected people, Ocimum gratissimum uses against various aliments has gained widespread acceptance in developing as well as developed nations (kolewale et al,. 2011). Diabetes mellitus is an important chronic metabolic disorder of public health concern, it occurs either as a result of pancreatic defects in insulin secretion or the failure of the effectively utilize secreted insulin or both. Hyperglycemia is a common consequence of uncontrolled diabetes, which may over time lead to serious damage to vascular tissue, heart, eye nerve and kidney.

            Plants with known and suspected therapeutic potencies have been longed used in the alternative and complementary medicine. Numerous scientific reports exist, describing the relatively low toxicity and effectiveness in selected plants in management of diabetes.

            Numerous scientific report exists, descending the relatively low toxicity and effectiveness of selected plants in the management of diabetes. It is unfortunate that a large number of those studies were conducted using either normoglycermeia or animal models.

 Inducing of diabetes using either >100mg/kg body weight of Alloxan into these animal models, with recent review showing that Alloxan was one of the most frequently used diabetogenic agent globally.

 High dose of the chemical are cytotoxic to the pancreatic beta cells giving rise to insulin deficiency.

    1.  STATEMENT OF THE PROBLEM

The statement of the problem in this research work is that the efficacy of Ocimum gratissimum leaf extract through assaying of biochemical parameters of alloxan-induced diabetic rats.

VISION: To determine the effect Ocimum gratissimum methanol leaf extract has alloxan-induced diabetic rats through assaying of biochemical parameters.

ISSUE STATEMENT: Diabetes mellitus has become one of the prevailing health cases people face today, with no or little knowledge on how it can be controlled and managed, it could worsen beyond control and as far as oxidative damage is concerned, it would multiple the generation of free radicals.

METHODS: Apart from modern drugs that can be used to control and manage diabetes, the use of Ocimum gratissimum menthol leaf extract was employed in other to determine the effect on alloxan-induced diabetic rats through assaying of biochemical parameter.

1.3 AIMS AND OBJECTIVE

1.3.1 AIM OF THE STUDY

The aim of this study is to determine the effects of the methanol leaf extract of Ocimum gratissimum on the oxidative and biochemical parameter of alloxan –induced diabetic rats.

      1. OBJECTIVES OF THE STUDY

Specifically, the study sort to:

  1. Determine the effects of the extract on MDA level
  2. Determine the effects of the extract on lipid profile
  3. Determine the effects of the extract on antioxidant enzyme activity.

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