EFFECTS OF GONGRONEMA LATIFOLIA AND OCIMUM GRATISSIMUM COMBINED AQUEOUS EXTRACTS ON ALLOXAN-INDUCED DIABETIC RATTUS NOVERGICUS

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ABSTRACT

The effect of combined aqueous leaves extracts of O. gratissimum and G. latifolium on alloxan-induced diabetic Rattusnorvegicuswas investigated for 28 days. Lethal dose (LD50) and phytochemical compositions were determined by standard methods. A total of 90 rats were divided into six groups (1, 2, 3, 4, 5, 6) of fifteen rats per group, with each group comprising of three replicates of five rats each. The treatment group 2, 3 and 4 were administered 100 mg/kg, 250 mg/kg and 350 mg/kg respectively. Group 1(normal control), 5 (standard control) and 6 (negative control) were administered distilled water, standard drug (Glibenclamide) and distilled water respectively. All the administration was done by oral intubation. Body weight, blood glucose levels, haematological, lipid profiles, biochemical parameters and histopathology were determined using standard methods. A non-significant decrease (p> 0.05) was recorded in the body weight of rats. Significant decreases (p< 0.05) were recorded in the blood glucose levels of rats. There were no significant differences (p> 0.05) in the lipid profiles and liver enzymes. The effects of O. gratissimum and G. latifolium combined extracts on other haematological parameters were not significant (p> 0.05) but a significant increase (p< 0.05) was recorded in the haemoglobin concentration. No significant (p> 0.05) histopathological lesions were observed in the liver while significant pathologic renal degeneration (p < 0.05) was observed in kidney histology. The result therefore, suggests that G. latifolium and O. gratissimum have hypoglycaemic, non- haematotoxic and hepatoprotective effects on alloxan–induced diabetic rats. Generally, they are non-toxic even at high dose of 350mg/kg body weight.

TABLE OF CONTENTS

Title page
Table of Contents
Abstract

CHAPTER ONE: INTRODUCTION AND LITERATURE REVIEW
1.1       Introduction
1.1.1    Justification of the Study
1.1.2    Objectives of the Study
1.2       Literature Review
1.2.1    Ocimumgratissimum
1.2.1.1 Antidiabetic potentials of Ocimumgratissimum
1.2.1.2 Hepatoprotective potentials of Ocimumgratissimum
1.2.2    Gongronemalatifolium
1.2.2.1 Antidiabetic potentials of Gongronemalatifolium
1.2.2.2 Hepatoprotective Potentials of Gongronemalatifolium
1.2.2.3 Effects of Gongronemalatifolium on haematological parameters
1.2.3    Diabetes
1.2.3.1 Diabetes insipidus
1.2.3.2 Diabetes mellitus
1.2.3.3 Types of diabetes mellitus
1.2.4    Hyperglycaemia and hypoglycaemia
1.2.5    Causes of diabetes
1.2.6    Symptoms of diabetes mellitus
1.2.7    Diagnosis, treatment and management of diabetes mellitus
1.2.8    Alloxan
1.2.9    Pancreas
1.2.10 Insulin

CHAPTER TWO: MATERIALS AND METHODS
2.1       Plant Materials
2.2       Preparation of Plant Material
2.3       Animal Model
2.4.      Induction of Diabetes Mellitus
2.5       Phytochemical Analysis
2.5.1    Alkaloid determination
2.5.2    Determination of total phenols
2.5.3    Determination of tannin content
2.5.4    Saponin determination
2.5.5    Flavonoid etermination
2.6       Lethal Dose (LD50) Determination
2.7       Experimental Design
2.8       Determination of Blood Glucose Concentration
2.9       Haematological Analysis
2.9.1    Haemoglobin estimation
2.9.2    Red blood cell count
2.9.3    Haematocrit
2.9.4    Erythrocyte sedimentation rate
2.9.5    Red blood cell indices
2.9.6    White blood cell count
2.10     Lipid Profile Analysis
2.10.1 Total cholesterol
2.10.2 Lipoprotein assay
2.10.2.1 High density lipoprotein cholesterol
2.10.2.2Low density lipoprotein
2.10.3 Triacylglycerol
2.11     Biochemical Analysis
2.11.1 Assay for alanine aminotransferase (ALT)
2.11.2 Assay for alkaline phosphatase (ALP)
2.11.3 Assay for aspartate aminotrasferase (AST)
2.12     Histopathology
2.13     Statistical Analysis

CHAPTER THREE: RESULTS
3.1       Phytochemical compositions of the aqueous leaves extracts of Ocimumgratissimum and Gongronemalatifolia
3.2       Lethal dose (LD50) of the mixed aqueous leaves extracts of Ocimumgratissimum and Gongronemalatifolia
3.3       Effects of the aqueous extracts of the combined plant extracts on the body weight of rats
3.4       Effects of the aqueous extracts of the combined plant extracts on the blood glucose concentration (mg/dL)
3.5       Effects of the aqueous extracts of the combined plant extracts on the total white blood cell count
3.6       Effects of the aqueous extracts of the combined plant extracts on the haemoglobin concentration
3.7       Effects of the aqueous extracts of the combined plant extracts on the total red blood cell count (×1012/L)
3.8       Effects of the aqueous extracts of the combined plant extracts on the haematocrit (HCT)(%)
3.9       Effects of the aqueous extracts of the combined plant extracts on mean corpuscular volume (MCV) (fL)
3.10     Effects of the aqueous extracts of the combined plant extracts on the mean corpuscular haemoglobin (pg)
3.11     Effects of the aqueous extracts of the combined plant extracts on the mean corpuscular haemoglobin concentration (g/dL)
3.12     Effects of the aqueous extracts of the combined plant extracts on the red cell distribution width- coefficient of variation (%)
3.13     Effects of the aqueous extracts of the combined plant extracts on the red cell distribution width – standard deviation (fL)
3.14     Effects of the aqueous extracts of the combined plant extracts on the total platelet count (× 109)
3.15     Effects of the aqueous extracts of the combined plant extracts on the mean platelet volume (fL)
3.16     Effects of the aqueous extracts of the combined plant extracts on the platelet distribution width
3.17     Effects of the aqueous extracts of the combined plant extracts on the plateletcrit (%)
3.18     Effects of the aqueous extracts of the combined plant extracts on the aspartate aminotransferase concentration (mg/dL)
3.19     Effects of the aqueous extracts of the combined plant extracts on the alanine transferase concentration (mg/dL)
3.20     Effects of the aqueous extracts of the combined plant extracts on the alkaline phosphatase concentration (mg/dL)
3.21     Effects of the aqueous extracts of the combined plant extracts on the total cholesterol concentration (mg/dL)
3.22     Effects of the aqueous extracts of the combined plant extracts on the triglyceride concentration (mg/dL)
3.23     Effects of the aqueous extracts of the combined plant extracts on the high density lipoprotein concentration (mg/dL)
3.24     Effects of the aqueous extracts of the combined plant extracts on the low density lipoprotein concentration (mg/dL)
3.25     Histopathological Effects of the Mixed Aqueous Leaves Extracts of Ocimumgratissimum and Gongronemalatifolia on Some Organs of Experimental Rats
3.25.1  Effects of the mixed aqueous leaves extracts of Ocimumgratissimum and Gongronemalatifolia on the liver histology of experimental rats
3.25.2 Effects of the mixed aqueous leaves extracts of Ocimumgratissimum and Gongronemalatifolia on the kidney histology of experimental rats

CHAPTER FOUR: DISCUSSION AND CONCLUSION
4.1       Discussion
4.2       Conclusion
References


CHAPTER ONE


INTRODUCTION AND LITERATURE REVIEW

1.2        Introduction

Diabetes mellitus is a prevalent disease affecting the citizens of both developed and developing countries. It is estimated that 25% of the world population is affected by this disease (Maiti et al., 2004). It is a chronic disease characterized by elevated blood glucose levels and disturbances in carbohydrate, fat and protein metabolism. Diabetes mellitus results when pancreatic beta cells are unable to maintain adequate insulin secretion to prevent hyperglycaemia. It is observed as the body’s inability to effectively regulate the sugar balance which leads to severe complications such as hyperglycaemia, obesity, neuropathy, nephropathy, retinopathy, cardiopathy, osteoporosis and coma leading to death. Pancreatic damage resulting in the dysfunction of α and β cells causes disordered glucose homeostasis. In diabetic individuals, the regulation of glucose levels by insulin is defective, either due to defective insulin production which is called insulin-dependent diabetes mellitus (IDDM, Type 1) or due to insulin resistance that is termed non insulin-dependent diabetes mellitus (NIDDM, Type 2) (Vijan, 2010).
Type 1 diabetes usually begins in childhood and is thought to be a result of autoimmune destruction of the pancreatic beta cells (the cells that produce insulin; also called islet cells). Destruction of the beta cells results in a complete loss of insulin production, thereby necessitating insulin injections to maintain blood sugar control. Type 2 diabetes is usually diagnosed after 40 years of age. Type 2 diabetes is frequently associated with insulin resistance and normal or even elevated levels of insulin, although subnormal insulin levels are also seen in some type 2 diabetics.
Gestational diabetes is characterized by hyperglycemia during pregnancy and usually disappears after the child is delivered. However, even though gestational diabetes may be relatively short-lived, it can compromise the health of both mother and foetus. Diabetes is associated with a number of significant medical problems. Severe hyperglycemia may result in coma or even death. Milder hyperglycemia, if present for many years, increases the risk of cardiovascular disease, which can manifest as a heart attack, congestive heart failure, stroke, gangrene of the extremities (necessitating amputation in some cases), or kidney failure (HMW, 2004).

The normal range for blood glucose level is between 70 – 110 mg/dl. Insulin is a hormone that helps to maintain normal blood glucose level by making the body’s cell absorbs glucose (sugar) so that it can be a source of energy. In people with diabetes, glucose levels....


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