DIETARY MANAGEMENT OF DIABETES WITH UNRIPE PLANTAIN (MUSA PARADISIACA): ITS EFFECTS ON GLUT1 AND GLUT4 GLUCOSE TRANSPORTERS AND OTHER BIOCHEMICAL PARAMETERS IN STREPTOZOTOCIN-INDUCED DIABETIC RATS

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ABSTRACT

This study was aimed at investigating the biochemical basis of dietary management of diabetes mellitus using unripe plantain in streptozotocin (STZ) induced diabetic rats. The effects of unripe plantain incorporated feeds on the regulation of glucose transport protein 1 (GLUT 1), glucose transport protein 4 (GLUT 4) and AMP-activated protein kinase (AMPK) activities as well as body weights, glycated hemoglobin, total protein, lipoprotein lipase, intestinal amylase and fasting blood glucose (FBG) levels were determined using appropriate biochemical methods. Twenty five (25) male albino wistar rats were divided into five (5) groups of five (5) rats each. Group 1 (non-diabetic), group 2 (diabetic, not treated) and group 3 (diabetic, treated with glibenclamide 5mg/kg body weight) all received standard rat feed. Group 4 (diabetic, fed unripe plantain incorporated feed 600mg/kg body weight) and group 5 (diabetic, fed unripe plantain incorporated feed 800mg/kg body weight). The experiment lasted for 28days. The fasting blood glucose levels and body weights were measured weekly, whereas all other biochemical parameters were measured at the end of the experiment. There was no significant (P>0.05) difference in the body weights of rats in groups 3 and 5 when compared to group 1 rats, whereas groups 4 and 2 rats showed significant (P<0.05) increase and decrease respectively. There was no significant (P>0.05) difference in FBG levels of groups 3, 4 and 5 rats when compared to group 1 rats, however, group 2 rats showed a significant (P<0.05) increase in FBG levels. Groups 2, 3 and 4 rats showed significant (P<0.05) increase in glycated hemoglobin levels whereas group 4 rats showed a non significant (P>0.05) increase when compared to rats in group 1. All the other groups showed significant (P<0.05) decrease in total protein when compared to group 1 rats. There was no significant (P>0.05) difference in lipoprotein lipase activity of groups 3, 4 and 5 rats while rats of group 2 showed a significant (P<0.05) decrease when compared to group 1 rats. Groups 3, 4 and 5 rats showed a significant (P<0.05) increase in GLUT 4 levels whereas group 2 rats showed a significant (P<0.05) decrease when compared to group 1 rats. Rats in group 4 and group 5 showed a non significant (P>0.05) difference whereas rats in group 2 and group 3 showed a significant decrease in GLUT 1 levels when compared to group 1 rats. There was no significant (P>0.05) difference in AMPK levels of all the groups. The results obtained show that unripe plantain possesses anti-diabetic properties and may be effective as a dietary management option for diabetics.





TABLE OF CONTENTS

Title Page                                                                                                                    i

Declaration                                                                                                                  ii

Certification                                                                                                                iii

Dedication                                                                                                                  iv

Acknowledgments                                                                                                      v

Table of Contents                                                                                                       vi

List of Tables                                                                                                              x

List of Figures                                                                                                             xi

Abstract                                                                                                                      xii

 

CHAPTER 1: INTRODUCTION

1.1              Definition Of Diabetes Mellitus (DM)                                                           1

1.2       Complications of Diabetes Mellitus                                                                2

1.3       Effect of Diabetes Mellitus on Glucose Transporters                                     2

1.4       Prevalence of Diabetes Mellitus in the World                                                3

1.5       Orthodox medicines Used in Management of Diabetes                                 4

1.5.1    Limitations of orthodox medicines in management of diabetes mellitus       5

1.6       Therapeutic Plants Employed in the Management of Diabetes Mellitus        5

1.6.1    Challenges linked to the use of medicinal plants                                          7

1.7       Dietary Management of Diabetes Mellitus                                                     8

1.7.1    Use of unripe plantain                                                                                   8

1.8       Aim of the Study                                                                                            9

1.9       Objectives of the Study                                                                                  10

1.9.1   Specific objectives of the study                                                                       10

1.10     Justification of the Study                                                                               11

 CHAPTER 2: LITERATURE REVIEW

2.1       Definition of Diabetes Mellitus                                                                      12

2.1.1    Classification of diabetes mellitus                                                                  13

2.1.1.1 Type 1 diabetes mellitus                                                                                 13

2.1.1.2 Type 2 diabetes mellitus                                                                                 14

2.1.1.3 Gestational diabetes mellitus (GDM)                                                             15

2.2       Glucose metabolism                                                                                        16

2.2.1    Hormonal regulation of glucose metabolism                                                  17

2.3       Glucose Transport in Diabetes Mellitus                                                          17

2.3.1    GLUT4 glucose transporters                                                                           18

2.3.2    GLUT1 glucose transporters                                                                           20

2.3.3    Regulation of glucose transport                                                                      21

2.3.4    Disorders in glucose transport in diabetes                                                      21

2.3.5    Problems of glucose transport in diabetes mellitus                                         22

2.4       Roles of AMP-kinase in the Regulation of Glucose Transport                      24

2.4.1    Roles of AMPK in glucose metabolism                                                          25

2.5       Roles of Intestinal Amylase and Lipoprotein Lipase in Carbohydrate

            Metabolism                                                                                                     26

2.5.1    Intestinal amylase in diabetes                                                                         27

2.5.2    Lipoprotein lipase in diabetes                                                                         27

2.6       Complications of Diabetes                                                                              28

2.6.1    Nephropathy                                                                                                   29

2.6.2    Retinopathy                                                                                                    30

2.6.3    Neuropathy                                                                                                     31

2.6.4    Cardiovascular diseases                                                                                  32    2.7           Management of Diabetes                                                                              33

2.7.1    Exercise and diet                                                                                            33

2.7.2    Drugs                                                                                                              34

2.8       Induction of Diabetes                                                                                     35

2.8.1    Streptozotocin                                                                                                 35

2.8.2    Handling of streptozotocin during induction                                                 37

 

CHAPTER 3: MATERIALS AND METHODS                                                  

3.1       Materials                                                                                                         38

3.2       Reagents and Chemicals                                                                                 38

3.3       Collection and Preparation of Plant                                                                39

3.4       Formulation of Test Feed                                                                               39

3.5       Animal Experiments                                                                                       40

3.5.1    Selection of animals                                                                                        40

3.5.2    Animal grouping                                                                                             40

3.6       Induction of Diabetes Mellitus                                                                       41

3.7       Experimental Procedure                                                                                  41

3.7.1    Assessment of body weights of the rats                                                         42

3.7.2    Determination of fasting blood glucose concentration                                   42

3.7.3    Assessment of feed intake                                                                              42

3.7.4    Collection of blood                                                                                         43

3.7.5    Determination of glycated hemoglobin                                                          43

3.7.6    Determination of serum lipoprotein lipase activity                                         44

3.7.7    Determination of intestinal amylase activty                                                   45

3.7.8    Preparation of tissue homogenate for GLUT1, GLUT4 and AMP-kinase

            analysis                                                                                                            46

3.7.9    Determination of glucose transporter 4 (GLUT4) concentration                   46

3.7.10  Determination of glucose transporter 1 (GLUT1) concentration                   47

3.7.11  Determination of AMP-kinase concentration                                                 48

3.7.12  Determination of serum total protein                                                              48

3.8       Statistical Analysis                                                                                          49

 

CHAPTER 4: RESULTS AND DISCUSSION

4.1       Results                                                                                                            50

4.1.1    Effect of unripe plantain incorporated feed on body weights of treated

            rats                                                                                                                  50

4.1.2    Effect of unripe plantain incorporated feed on fasting blood glucose

            levels of treated rats                                                                                        51

4.1.3    Effect of unripe plantain incorporated feed on feed intake                           52

4.1.4    Effect of unripe plantain incorporated feed on glycated hemoglobin

            levels                                                                                                               53

4.1.5    Effect of unripe plantain incorporated feed on serum lipoprotein lipase

            activity                                                                                                                        54

4.1.6    Effect of unripe plantain incorporated feed on intestinal amylase

            activity                                                                                                                        55

4.1.7    Effect of unripe plantain incorporated feed on GLUT 4 concentration         56

4.1.8    Effect of unripe plantain incorporated feed on GLUT 1 concentration         57

4.1.9    Effect of unripe plantain incorporated feed on levels of AMP-kinase           58

4.1.10  Effect of unripe plantain incorporated feed on levels of serum total

            protein                                                                                                             59

4.1.11  Proximate composition of test and animal feeds                                            60

4.2       Discussion                                                                                                       61

 

CHAPTER 5: CONCLUSION

5.1       Conclusion                                                                                                      66

            References                                                                                                      67

            Appendices                                                                                                     79

 

 

 

 

 

LIST OF TABLES

                                                                                                                                   PAGE

3.1       Composition of the test feeds                                                                        40

4.1       Body weights of treated rats                                                                          50

4.2       Fasting blood glucose levels of treated rats                                                    51

4.3       Feed intake of treated rats                                                                              52

4.4       Glycated hemoglobin levels of treated rats                                                    53

4.5       Lipoprotein lipase activity of treated rats                                                       54

4.6       Intestinal amylase activity of treated rats                                                       55

4.7       GLUT 4 concentration of treated rats                                                                        56

4.8       GLUT 1 concentration of treated rats                                                                        57

4.9       AMP-Kinase levels of treated rats                                                                  58

4.10     Serum total protein levels of treated rats                                                        59

4.11     Proximate composition of test and animal feeds                                            60

 

 

 


 

 

 

LIST OF FIGURES

                                                                                                                                  PAGE

2.1       Structure of the Insulin-Regulated GLUT4 Glucose Transporter

Protein                                                                                                             19

2.0       Structure of streptozotocin                                                                             35

 

 

 

1

 

 

CHAPTER 1

 

INTRODUCTION

 

 

1.1              DEFINITION OF DIABETES MELLITUS (DM)

 

Diabetes Mellitus is a global metabolic disease characterized by hyperglycaemia and affects essential biochemical pathways in the body resulting to complications such as; renal disorders, neuropathy, retinopathy, ketoacidosis, and cardiovascular diseases. It is caused by impaired regulation of the metabolism of carbohydrate and lipids where although there is sufficient supply of glucose, the body still behaves as if it is starved; hence glucose produced by the liver is not utilized by other tissues (Horton et al., 2006). Its symptoms include an upsurge in the need to urinate, raised thirst, and a surge in feelings of hunger (Cooke and Plotnick, 2008). It is classified into three types: Type 1 diabetes mellitus which can be distinguished by damage to the insulin-producing beta-cells of the pancreatic islets of Langerhans, which results in insulin deficiency. Type 1 diabetes is divided into two types namely immune-mediated or idiopathic. The majority of type 1 diabetes results from immune-mediated type, whereby a  T-cell-mediated  autoimmune attack results in damage to the beta-cells and consequently insulin (Rother, 2007). It occurs at an early age though it also affects adults. Type 2 diabetes mellitus is typified by gradual development of resistance to insulin and beta-cell dysfunction, in addition to decreased insulin secretion in some cases (Zimmet et al., 2001; Gardner, 2011). The inability of body tissues to act in response insulin appears to result from the insulin receptor. The risk for type 2 diabetes rises with an increase in age and body weight as well as a sedentary lifestyle (Zimmet, 1992). It is the most occurring of diabetes and usually occurs in adults. Gestational diabetes is the third principal type and arises after pregnant women who have not had diabetes before develop high blood-sugar levels (WHO, 2013). This type of diabetes typically fades away once the child is born.

 

 

1.2              COMPLICATIONS OF DIABETES MELLITUS

 

The problems identifiable with diabetes mellitus comprise retinopathy, nephropathy, and neuropathy. Diabetic patients suffering from any kind diabetes for some time are susceptible to the above-mentioned problems (Nathan, 1993).

 

 

1.3  EFFECT OF DIABETES MELLITUS ON GLUCOSE TRANSPORTERS

 

Glucose movement across the cell membrane is the rate limiting step of glucose utilization and is regulated by a family of membrane proteins known as Glucose Transporters (GLUTs) (Mueckler and Thorens, 2013). Notwithstanding whether there is translation or transcription, insulin and also physical activity causes acute stimulation and recruitment of GLUT4 to the surfaces of the cells of skeletal muscle and fat tissues (Herman and Kahn, 2006). Insulin also stimulates GLUT1 which is expressed in the blood. Insulin appears to be the main hormone affected in diabetes since the pancreas is involved in insulin production. Ryder et al in 2000 showed that GLUT4 translocation, in response to insulin, in the skeletal muscle of people suffering from type 2 diabetes is usually decreased by approximately 90%. The main cellular mechanism for clearance of glucose taken from foods is insulin-mediated glucose transport into the cells of skeletal muscle. Following the transport step, skeletal muscle accumulates glucose in the form of glycogen and breaks it down to produce energy when required. The main glucose transport protein which facilitates the movement of glucose into cells is GLUT4 a member of the sugar transport proteins family that contain 12-transmembrane domains. The GLUT4 glucose transporter is an important facilitator of glucose clearance out of the blood and is therefore a core controller of glucose homeostasis (Huang and Czech, 2007). Studies have proven that although glucose transporters move from their intracellular membrane compartment to the surface of the cells in response to insulin, the affinity of the transporters for glucose binding is not affected by insulin (Cushman and Wardzala, 1980; Suzuki and Kono, 1980). There is a reduced GLUT4 protein content in intracellular and plasma membranes of skeletal muscles of STZ-induced diabetic rats. In STZ-treated rats, insulin caused the mobilization of GLUT4 out of the vesicles, but the fusion of GLUT4 with the plasma membrane was diminished. Conversely, the GLUT1 transporter protein increased in the cell membrane of the diabetic rats (Klip et al.,1992).


 

1.4              PREVALENCE OF DIABETES MELLITUS IN THE WORLD

 

The World Health Organization estimates that, globally, 422 million adults aged over 18 years were living with diabetes in 2014 (WHO, 2016). South-East Asia and Western Pacific Regions have the highest occurrence of diabetes as estimated for the WHO, thus, this accounts for about half the cases of diabetes on earth. The people with diabetes (defined in surveys as those with a fasting blood glucose value of ≥7.0mmol/L or on medication for diabetes/hyperglycaemia) progressively rose within the last decades following growth in population, rise in the normal age of people in the population, and the rise in prevalence of diabetes at each age. Worldwide, between 1980 and 2014, people with diabetes have increased tremendously from 108 million to current numbers that are around four times higher. Forty percent of this rise is thought to be from increase in population, twenty eight percent from an increase in age, and thirty two percent from both increase in population and age (WHO, 2016). The occurrence of diabetes around the universe has increased from 4.7 percent to 8.5 percent between 1980 and 2014. This implies that occurrence has either been on the increase or has remained unchanged in every country (WHO, 2016). In the last decade, the incidence of diabetes has increased in both developing and under-developed countries than in high income countries (WHO, 2016). There are about 22 million diabetics resident in sub-Saharan Africa only. Nigeria, the most populated country in Africa has about 4 million diabetics out of the entire diabetics in sub-Saharan Africa. The problem with the issue of diabetes in Nigeria is that a significant number of diabetic cases are yet to be diagnosed and hence remain untreated (about 70%-80% of the 4 million) (Fasanmade and Dagogo-Jack, 2015).

 

Nigeria has a high occurrence of diabetes across the country. In the non-urban areas, diabetes occurs in 0-2% of the population, but in the urban areas the numbers higher at 5-10% (Fasanmade and Dagogo-Jack, 2015). In Nigeria, diabetes is uncommon in children, but reports from local medical facilities posit that there is a gradual increase in number of adolescents and children suffering from diabetes (Fasanmade and Dagogo-Jack, 2015).


 

1.5              ORTHODOX MEDICINES USED IN MANAGEMENT OF DIABETES

 

The therapy for type-1 diabetics is administration of exogenous insulin. Some type-2 diabetics also require exogenous insulin if they do not respond satisfactorily to oral hypoglycaemic drugs. Drugs used for the treatment of diabetes can be grouped into three. Drugs in group 1 boost the amount of endogenous insulin. They include the sulphonylureas which comprise the glinides, glibenclamide, glucagon-like peptide 1 (GLP-1) agonists, insulin analogs, and dipeptidyl peptidase-IV (DPP-IV) inhibitors. The glibenclamide and the glinides enhance insulin secretion by acting on the sulphonylurea receptor found in the pancreas. Dipeptidyl peptidase-IV (DPP-IV) inhibitors and glucagon-like peptide 1 (GLP-1) agonists effect their action on the ileal cells of the small intestine. Drugs in group 2 enhance insulin sensitivity. They include the thiazolidinediones, which are peroxisome proliferator-activated receptor gamma (PPARγ)agonists  and  metformin,  a  biguanide.  Thiazolidinediones  act  by reducing insulin resistance in the liver, fat cells and muscle while metformin inhibits gluconeogenesis in the liver. Drugs in group 3 comprise of α-glucosidase inhibitors like acarbose that lowers the breakdown of carbohydrates hence their bioavailability (Chehade and Mooradian, 2000).


 

1.5.1        Limitations of orthodox medicines in management of diabetes mellitus

 

All known treatment have shown inadequate effectiveness, tolerance and adverse effects based on their mode of action  (Moller, 2001; Rotenstein et al., 2012). One of the adverse effects of sulphonylureas is hypoglycaemia which if not well managed, leads to death (Erasmus et al., 1999). Thus the use of these drugs for the treatment of diabetes has resulted in severe adverse effects. Ongoing researches aimed at developing drugs with lower adverse effects have yielded little success (Levey, 2002). Notwithstanding the existing treatments available, attaining optimum glucose control continues to elude diabetics because of retrogression in β-cell function  (Wallace and Matthews, 2000). There is also increase in complications from diabetes and hyperglycaemic emergencies (Gill et al., 2009; Ogbera et al., 2007; Ogbera et al., 2009). In the light of these, the volume of drugs administered has risen to tremendously (Enwere et al., 2006). The high cost of these drugs and the need to take them continuously, even with the associated adverse effects has been identified as the reason why diabetics do not adhere to treatment schedule. Besides the above mentioned, there is the problem of inadequate accessibility to these orthodox medicine to treat diabetes in middle and low income countries.


 

1.6  THERAPEUTIC PLANTS EMPLOYED IN THE MANAGEMENT OF DIABETES MELLITUS


Following the side effects and high cost of diabetic drugs, patients usually turn to other forms of therapy like the medicinal plants (Yusuf et al., 2008). Many plant remedies known in folkloric medicine are being employed in treatment of hyperglycemia in diabetics. Some of these plant preparations have been substantiated by research studies and found to exert biological actions against the complications of diabetes and diabetes itself (Ezuruike and Prieto, 2014). Several mechanisms have been adduced for the hypoglycaemic action of medicinal plants. These include reabsorption of glucose by the kidneys, improved insulin secretion by beta-cells, improved glucose uptake by tissues following improved insulin sensitivity, rejuvenation/repair of beta cells, accelaration of glycogenesis and hepatic glycolysis, rise in size and quantity of cells of the islets of Langerhans, protective effects on the destruction of the beta-cells and/or prevention of oxidative stress that is possibly involved in damage to pancreatic beta-calls (Ismail-Beigi, 1993). In Nigeria, it is common to use plants (herbs or food plants) in isolation or along with medicines in diabetes management. The diets/medicinal plants frequently employed in treatment of diabetes in Nigeria include: sweet potato (Ipomoea batatas), acha (Digitaria exillis), okro (Abelmoschus esculentus), breadfruit (Treculia africana), bitterleaf (Vernonia amygdalina), beans (Phaseolus vulgaris), Aloe vera, garlic (Allium sativum), Momordica charantia,alligator pepper (Aframomum melegueta), (Eleazu and Okafor, 2012; Ezuruike and Prieto, 2014; Nwaoguikpe, 2010). The hypoglycaemic effect of these plants stems from the presence of certain biochemicals present in them. Lupenyl cinnamate, lupenyl - and cetate-amyrin β cinnamates and isolated α from the combined root and stems of locally obtained samples of Gongronema latifolium, otherwise known-masro’‘utazi’inIgbor and‘maduYoruba is commonly used as a food vegetable and is widely recognized for its traditional use in diabetes management) have recently been identified as the bioactive compounds, possessing both anti-hyperglycaemic effects in glucose-fasted rats in addition to its ability to stimulate insulin in INS-1 cells (Adebajo et al., 2013). Kolaviron is a biflavonoid isolated from extracts of the edible nuts of Garcinia kola Heckel (bitter kola), which is of significant importance in many regions of West Africa. It reduced blood glucose concentration in non-diabetic and alloxan-induced diabetic mice at a dose of 100 mg/kg, besides inhibiting rat lens aldose reductase (RLAR) activity (Iwu et al., 1990).  Some methoxy phenyl derivatives present in the rhizomes of Zingiber officinale Roscoe have been identified as aldose reductase inhibitors. They repress accumulation of sorbitol in human red blood cells and also reduce accumulation of lens galactitol in 30% of galactose-fed rats (Kato et al., 2006). Fractionating the bark of Cassia fistula L. showed that the plant contains catechin as its active compound. Catechin reduced blood glucose levels in STZ-induced diabetic rats by acting directly on enzymes involved in glucose breakdown and also on GLUT4 expression  (Daisy et al., 2010). Quercetin and epicatechin are both able to inhibit the development of advanced glycated end products (AGEs) as well as other complications of diabetes that result from oxidative stress conditions like erectile dysfunction, artherosclerosis, neuropathy, nephropathy and retinopathy (Rahimi et al., 2005). Bush mango (Irvingia gabonensis), mahogany (Khaya senegalensis), mango (Mangifera indica), and scent leaf (Ocimum gratissimum) all contribute to an all round treatment of diabetes as well as the prevention of diabetic complications because they contain quercetin, epicatechin and other effective flavonoids (Ezuruike and Prieto, 2014).


 

1.6.1        Challenges linked to the use of medicinal plants

 

The major challenges of traditional medicine are overall quality, safety and effectiveness of the medicines. Preservation and dosage measurements of the medicines are serious problems too. However, despite their challenges, herbal medicines afford research in science great prospect for development (Erah, 2002).


 

1.7              DIETARY MANAGEMENT OF DIABETES MELLITUS

 

Before the 1920s when insulin was introduced, the sole treatment for diabetes was dietary (Chandalia et al., 2000). After the coming of insulin and other drugs there exist quite a number of limitations in their use even though they have been useful for managing the disease. These limitations include their side effects and high cost as well as lack of accessibility to low income countries. Another important limitation is the need to combine two or more of these drugs for better effects. These reasons have made it preferable to embrace and adopt alternatives which include dietary management. Chandalia et al., 2000 showed that increasing consumption of soluble fiber is one effective dietary treatment. Its benefits in terms of serum cholesterol concentrations and colonic function are well established (Bruce et al., 2000). Another concept is the incorporation of foods with low glycaemic index in the diet. Miller, 1994 determined that diets that incorporate low glycaemic index foods reduce hyperglycemia in type 2 diabetes. In addition, the production of free fatty acids is usually lower after a meal that has a low glycaemic index. This is good for diabetics since insulin resistance is promoted by fatty acids. Lower insulin resistance after a meal translates into lower blood glucose concentrations. Results from various studies indicate that dietary manipulation produces dividends in diabetics. Combining a high fiber diet in addition to other foods with low glycaemic index successfully lowers blood glucose levels synergistically (Chandalia et al., 2000; Bruce et al., 2000). It is necessary to note that although carbohydrates that have low glycaemic index appear to be desirable, elimination of carbohydrates is not beneficial.


 

1.7.1        Use of unripe plantain

 

Plantain (Musa paradisiaca) is a major food crop in the tropical and sub-tropical regions of Africa and the staple supply of energy for many people dwelling in these regions (Adepoju et al., 2012). In Nigeria, roughly 2.4 million metric tonnes of plantain is produced from a state in the southern region (Folayan and Bifarin, 2011). In Nigeria, plantain is widely used in an array of ways and hence it is placed among the staple foods in this country. Musa paradisiaca (plantain) is a member of the Musaceae family, the perennial tropical giant plant. Plantain fruits can be consumed in any form (Nwokocha and William, 2009). It has been shown that unripe plantain contains 83.40% carbohydrate, 8.10% protein, 1.10% fat, 1.50% fiber, 0.46% moisture, and 1.10% ash. It is also abundant in vitamin K,  provitamin A, and  vitamin C. Saponins, alkaloids, tannins and flavonoids are also present in substantial amounts (Eleazu et al., 2011). Other studies have recorded the presence of  vitamin C, vitamin A, vitamin B complex, vitamin E, as well as high potassium in addition to low sodium in plantains (Marriott and Robinson, 1981). Plantain is also known to be rich in iron, fibre, vitamins, minerals, and serotonin (Jimmy and Okon, 2012). Unripe plantain is employed in traditional medical practice for the management of diabetes, treatment of anemia and treatment of ulcers (Ekpo et al., 2011). In Nigeria, meals made using unripe plantain have been adduced to lower postprandial glucose levels in diabetics (Willet et al., 2002). The ability of unripe plantain to reduce the prevalence of diabetes has been reported in experimental animals. Previous studies show that unripe plantain can ameliorate glomerular complications, liver hypertrophy and kidney hypertrophy in Streptozotocin-induced diabetic rats (Eleazu et al., 2010; Eleazu and Okafor, 2015). Its anti-hyperglycaemic effect has also been established (Iroaganachi et al., 2015).


 

1.8              AIM OF THE STUDY

 

The use of unripe plantain as a source of dietary management of diabetes has been established (Eleazu and Okafor, 2015; Iroaganachi et al., 2015). However, the mechanism by which unripe plantain exerts this function is not clear. Thus the possible effect of formulated unripe plantain diet in regulation of GLUT4 and GLUT1 (principal transporters of glucose) forms the basis of this work.

 

Based on above, the purpose of this study is to investigate the possible effect of formulated unripe plantain diet in the regulation of GLUT4 and GLUT1 transporters.


 

1.9              OBJECTIVES OF THE STUDY

 

The overall objective of this work is to determine the possible effect of a formulated unripe plantain diet on regulation of GLUT4 and GLUT1 and on other biochemical parameters such as AMPK, glycated haemoglobin levels, intestinal amylase, and lipoprotein lipase activity.


 

1.9.1        Specific objectives of the study

 

 

·         To investigate the possible effect of formulated meal of unripe plantain on the concentrations of GLUT4 and GLUT1 glucose transporters because a low concentration of the transporters indicates a low level or insensitivity of insulin.

 

·         To determine the AMPK activity as their effect is diminished in diabetic rats.

 

·         To investigate serum amylase and lipase activities as it indicates the existence of acute pancreatitis in the diabetic rats.

 

·         To ascertain the effect of the meal on body weights and feed intake because alteration in cellular metabolism manifest in body and organ dysfunction and weight alteration.

 

·         To determine the level of glycation of haemoglobin because levels of HbA1c is a very good indicator of diabetic state in rats.

 

·         To ascertain the effect of the meal on blood glucose levels because blood glucose levels are indicators of diabetes in the STZ-treated rats.

 

·         To determine the proximate analysis of the test feeds since components of food determine their impact on hyperglycemia



 

1.10          JUSTIFICATION OF THE STUDY

 

Following the alarming rate at which the incidence of diabetes is snowballing and in the light of the potentials that unripe plantain has in its management, there is need for studies that could help in our understanding of the biochemical basis for such dietary management of the disease. No doubt many studies have corroborated that unripe plantain is effective in diabetes management, this study however seeks to delve into the effect that unripe plantain has on regulation of GLUT4 and GLUT1 glucose transport proteins in the skeletal muscle and blood respectively in a bid to tackle hyperglycemia.

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