ATTENUATION OF HIGH-SUCROSE DIET-INDUCED METABOLIC SYNDROME IN ALBINO RATS BY METHANOL EXTRACT OF CURCUMA LONGA (TURMERIC)

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ABSTRACT


There is prevalence of Metabolic syndrome, a cluster of cardiometabolic symptoms including central obesity, hypertension, insulin resistance, chronic low-grade inflammation and dyslipideamia. The present study evaluated the possible attenuating role of low (500mg/kg/day) and high (1000mg/kg/day) doses of methanol extract of turmeric, a common food constituent, in high-sucrose diet-induced metabolic syndrome in rats. Forty two male Wistar rats were assigned to seven Groups of six rats each. Group 1 served as normal control and was given standard feed and distilled water only, Group 2 received standard feed and 500mg/kg/day of  turmeric extract, Group 3 received standard feed and 1000 mg/kg/day of turmeric extract, Group 4 served as high sucrose diet (HSD) control and received distilled water and a high sucrose diet containing 390g of powdered diet (standard diet), 200g of sweetened condensed milk, 140g of sucrose (table sugar), and 270mL of water per kilogram of diet, Group 5 received HSD and 500 mg/kg/day of turmeric extract, Group 6 received HSD and 1000 mg/kg/day of turmeric extract while Group 7 received HSD and 2mg/kg/day of the standard drug for 28days. The HSD induced body weight gain and significantly increased blood glucose levels in Group 4. However, body weights and blood glucose were significantly reduced in treated Groups compared to Group 4. There was also a significant (P˂0.05) increase in SOD, GSH and catalase in the groups treated compared to Group 4. Furthermore the results showed significant (P<0.05) decrease in serum lipid profiles concentrations (TG, TC, LDL and VLDL) in all treated Groups which increased significantly relative to the HSD control Group, while HDL was significantly  increased in all treated groups compared to Group 4. It was also observed from the result that the serum insulin levels were significantly increased in all treated groups compared to Group 4. Thus at the tested doses, turmeric attenuated the studied indicators of metabolic syndrome suggesting that its use could be beneficial in the management of obesity and related diseases associated with metabolic syndrome.

 

 

 

 

 

TABLE OF CONTENTS

Title Page                                                                                                              i

Declaration                                                                                                            ii

Certification                                                                                                          iii

Dedication                                                                                                            iv

Acknowledgements                                                                                              v

Table of contents                                                                                                  vi

List of Tables                                                                                                        xi

List of Figures                                                                                                       xii

List of Plates                                                                                                         xiv

Abstract                                                                                                                            xv

Chapter 1:      INTRODUCTION

1.0       Background of Study                                                                         1

1.1       Aim of study                                                                                       2

1.2       Objectives                                                                                           2

1.3       Justification                                                                                         2

Chapter 2:      LITERATURE REVEIW

2.1       Overview of Turmeric                                                                         4

2.2       Nutritional Value of Turmeric                                                5

2.3       Medicinal Uses of Turmeric                                                                6

2.3.1    Anti-Inflammatory properties                                                             6

2.3.2    Antioxidant properties                                                                        6

2.3.3    Hepatoprotective properties                                                                6

2.3.4    Anticarcinogenic properties                                                                7

2.3.5    Antidiabetic properties                                                                       10

2.3.6    Antimicrobial properties                                                                     10

2.3.7    Cardiovascular diseases                                                                      11

2.3.8    Gastrointestinal disorders                                                                   11

2.3.9    Dyspepsia and gastric ulcer                                                                12

2.3.10  Neurological disorders                                                                        12

2.4       Metabolic syndrome                                                                           13

2.5       Obesity and the Metabolic Syndrome                                                            16

2.6       Impaired Fasting Glucose                                                                   17       

2.7       Dyslipidemia                                                                                       17

2.8       Hypertension                                                                                       19

2.9       Insulin Resistance                                                                               19

2.10     Choice of A Suitable Extraction Solvent                                           21

2.11 Oxidative Stress Markers                                                                        21

2.11.1 Reduced glutathione (GSH)                                                                21

2.11.2 Glutathione peroxidase (GPx)                                                             22

2.11.3 Catalase                                                                                                22

2.11.4 Superoxide dismutase (SOD) activity                                                 22

2.11. 5 Thiobarbituric acid reactive substances (TBARS)                             23

2.12     Blood Lipid Profile                                                                             23

2.12.1  Cholesterol                                                                                          24

2.12.2  Very low-density lipoproteins (VLDL)                                              25

2.12.3  Low density lipoprotein (LDL)                                                          25

2.12.4  High density lipoprotein (HDL)                                                         25

2.12.5  Triacylglycerols                                                                                   26

2.13     C-peptide                                                                                            26

2.14     Hexokinase                                                                                         27

2.15     Glycogen                                                                                             27

Chapter 3:      MATERIALS AND METHODS

3.1       Animals                                                                                               28

3.2       Chemicals                                                                                           28

3.3       Plant material collection and processing                                             28

3.4       Methanol Extract Preparation                                                             28

3.5       Experimental design                                                                           29

3.6       Diets                                                                                                   29

3.7       Ethical Clearance/Approval                                                               30

3.8       Blood Collection and Separation                                                       30

3.9       Determination of Relative Organ Weights                                         30

3.10     Histological Study                                                                             30

3.11     Determination of biochemical parameters                                          32

3.11.1  Determination of Serum Insulin                                                         32

3.11.2  Biochemical Assay of Serum Glycogen                                             32

3.11.3  Biochemical assay of serum hexokinase activity                                33

3.11.4  Biochemical Assay of serum C-peptide activity                                 34

3.11.5  Estimation of blood glucose                                                               34

3.12 Determination of Lipid Profile                                                               34

3.12.1 Total Cholesterol concentration                                                           35

3.12.2 Triacylglycerol concentration                                                               35

3.12.3 High density lipoprotein cholesterol concentration                             35

3.12.4  Very low density lipoprotein cholesterol concentration                     35

3.12.5  Low density lipoprotein cholesterol concentration                             36

3.13     Assay of Superoxide Dismutase (SOD) Activity                               36

3.14     Assay of Catalase (CAT) Activity                                                      36

3.15     Assay of Glutathione Peroxidase (GPx) Activity                              37

3.16     Estimation of extent of lipid peroxidation (Malondialdehyde)

 Concentration                                                                                    37

3.17     Assay of reduced Gluthathione (GSH) Activity                                38

3.18     Measurement of Lee Obesity Index                                                   38

3.19     Statistical Analysis                                                                              38

 

Chapter 4:     RESULTS AND DISCUSSION     

            4.1       Results                                                                                                39        4.1.1    Liver Histology                                                                                57

            4.1.2    Pancreas Histology                                                                             59

            4.2       Discussion                                                                                           61

Chapter 5:      CONCLUSION AND RECOMMENDATIONS

            5.1       Conclusion                                                                                          66

            5.2       Recommendation                                                                                66

References                                                                                          67

Appendices                                                                                         76

 

 

 

 

 

 

LIST OF TABLES

                                                                                                                       

2.1:      The American Heart Association and the National Heart, Lung and Blood

Institute criteria for defining the metabolic syndrome.                                  15

4.1:      Body Weight gain/loss of the experimental rat groups over the experimental

 period                                                                                                             39

4.2:      Body Weight gain/loss of the experimental rat groups over the experimental

period                                                                                                              40

4.3:      Difference in fasting blood glucose concentrations at the beginning and

 end of the experimental period.                                                                     55

 

 

 

 

 

 

LIST OF FIGURES

 

2.1       Flow chart showing factors leading to the metabolic syndrome (MetS).       14

4. 1      Effect of Methanol extract of C. Longa on Total Cholesterol Concentration of the

 rat groups                                                                                                       41

4.2       Effect of Methanol extract of C. Longa on High-density lipoproteins

 Concentration of the rat groups                                                                     42

4.3       Effect of Methanol extract of C. Longa on Triacylglycerol Concentration of the

rat groups                                                                                                        43

4.4       Effect of Methanol extract of C. Longa on Low-density lipoproteins

Concentration of the rat groups                                                                      44

4.5       Effect of Methanol extract of C. Longa on Very-low-density lipoprotein

 Concentration of the rat groups                                                                     45

4. 6      Glutathione peroxidase (GPx ) concentration of the rat groups                    46

4. 7      Reduced Gluthathione (GSH) concentration of the rat groups                      47

4. 8      Superoxide dismutase (SOD) concentration of the rat groups                       48

4.9       Catalase (CAT) concentration of the rat groups                                             49

4.10     Thiobarbituric Acid Reactive Substances (TBARS) concentration of the

 rat groups                                                                                                       50

4. 11:   Serum Insulin Concentration of the rat groups                                               51

4. 12:   Glycogen Concentration of the rat groups                                                     52

4. 13:   C-peptide Concentration of the rat groups                                                    53

4. 14:   Hexokinase Concentration of the Concentration rat groups                         54

 

 

 

 

 

 

 

 

LIST OF PLATES

2.1       Turmeric Plant                                                                                                5

4.1       Photomicrograph of the liver section of rats in group 1 (Normal Control Group)    

      Mag x100.                                                                   57

4.2.      Photomicrograph of the liver section of rats in group 2, Mag x400.              57

4. 3.     Photomicrograph of the liver section of rats in group 3, Mag x100.             58

4.4.      Photomicrograph of the liver section of rats in group 4, Mag x100.              58

4. 5.     Photomicrograph of the liver section of rats in group 5, Mag x400.              58

4.6.      Photomicrograph of the liver section of rats in group 6, Mag x100.              58

4. 7.     Photomicrograph of the liver section of rats in group 7, Mag x100.              58

4.8.      Photomicrograph of the pancreas section of rats in group 1, Mag x400.       59

4. 9.     Photomicrograph of the pancreas section of rats in group 2, Mag x400.       59

4.10.    Photomicrograph of the pancreas section of rats in group 3, Mag x400.       60

4.11.    Photomicrograph of the pancreas section of rats in group 4, Mag x400.       60

4.12.    Photomicrograph of the pancreas section of rats in group 5, Mag x400.       60

4.13.    Photomicrograph of the pancreas section of rats in group 6, Mag x400.       60

4.14     Photomicrograph of the pancreas section of rats in group 7, Mag x400.       60

 

 

 

CHAPTER 1

INTRODUCTION

 

1.0       BACKGROUND OF STUDY

There has been a steady increase in the incidence of diseases associated with metabolic syndrome also known as syndrome X, insulin resistance syndrome, or dysmetabolic syndrome. Metabolic syndrome is the clustering of cardiometabolic symptoms including central obesity, hypertension, insulin resistance, chronic low-grade inflammation and dyslipideamia (Jaspinder, 2014).

Diabetes mellitus is a chronic metabolic disorder characterized by hyperglycemia resulting from defect in insulin secretion, insulin action or both. It is associated with high morbidity and mortality from long-term micro-vascular and macro-vascular complications (Jeevangi et al., 2013).

Curcuma longa Linn is also known as turmeric. The plant is a tropical perennial plant which belongs to the same family as ginger (Zingiberaceae), native to India and Indonesia and is cultivated in many parts of the world. It is otherwise called the golden spice of life" and is a standout amongst the most fundamental spices used throughout the world. Turmeric is the zest that gives curry its yellow shading. It has been utilized in India for many years as a zest and restorative herb. Turmeric is utilized in coloring industry to color garments and textiles. All inclusive, the world production of turmeric is between 10-15kg every year and out of these production figures, India represents over 78% of the yearly production for turmeric. India is trailed by China and Myanmar in Asia. Nigeria is the fourth biggest maker of turmeric with about 3% of the worldwide yearly production.

The active component of turmeric, curcumin, has caught attention as a potential treatment for diabetes and its complications primarily because it is a relatively safe and inexpensive agent that reduces glycemia and hyperlipidemia in rodent models of diabetes. (Zhang et al., 2013).


1.1       AIM OF STUDY

The aim of this study is to determine the possible attenuating role of orally administered turmeric grown in the rainforest zone of South-Eastern Nigeria on bio-indicators of metabolic syndrome in rats fed high sucrose diet.


1.2       OBJECTIVES

The objectives of this study were to:

1.                  Determine the effect of methanol extract of Curcuma longa on the oxidative stress parameters: Superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GSH), catalase and thiobarbituric acid reactive substances (TBARS) of high sucrose diet-induced metabolic syndrome in Wistar rats.

2.                  Investigate the fasting blood glucose, Serum insulin, glycogen, C-peptide, hexokinase and body weight in normal and high sucrose diet-induced metabolic syndrome in Wistar rats.

3.                  Determine the effect of methanol extract of Curcuma longa on the lipid profile (total cholesterol, high density lipoprotein cholesterol, triacylglycerol, very low density lipoprotein, and low density liopoprotein) concentrations of high sucrose diet-induced metabolic syndrome in Wistar rats.

4.                  Check the Lee’s obesity index diet-induced metabolic syndrome in rats and the attenuating effects of methanol extracts of Curcuma longa on it.

5.                  Examine the effects of methanol extracts of Curcuma longa on the histoology of the liver and pancreas of high sucrose diet-induced metabolic syndrome in rats.

 

1.3       JUSTIFICATION

Prevalence of metabolic syndrome is rapidly increasing worldwide without effective management regime. In addition, the currently available drugs have number of adverse effects necessitating increased search for indigenous plant – based remedy in line with the WHO recommendation sequel to beneficial uses of medicinal plants in the treatment of diabetes mellitus, a compound of metabolic syndrome. The reported availability of bioactive compounds in turmeric world over warranted this study on turmeric grown in South Eastern Nigeria.

 

 

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