Antidiarrhoeal Effect of Unripe Musa paradisiacae Pulp and Peel Homogenates on Castor Oil-Induced Diarrhoea in Wistar Albino Rats

Antidiarrhoeal Effect of Unripe Musa paradisiacae Pulp and Peel Homogenates on Castor Oil-Induced Diarrhoea in Wistar Albino Rats.

ABSTRACT

Musa paradisiacae commonly known as plantain is a  rhizomatous  perennial  crop used as a source of starchy staple for millions of people in Nigeria. Different parts of the plant have been used  in the treatment of various  ailments and there are claims that it has antidiarrhoeal activity.

This study is therefore aimed  at determining the  effects  of unripe Musa paradisiacae pulp and peel homogenates on castor oil-induced diarrhoea in Wistar albino rats.

The qualitative phytochemical constituents of Musa paradisiacae pulp and peel were found to be flavonoids, saponins, soluble carbohydrates, tannins, reducing sugars, hydrogen cyanide, steroids, alkaloids, and glycosides. The LD₅₀ results showed no toxicity up to 5000 mg/kg body weight.

Rats were divided into 7 groups of 4 rats each. The groups were pre-treated as follows:  group 1: normal saline (control); group 2: 3 mg/kg lomotil (standard drug); groups 3 and 4: 200 and 400 mg/kg unripe Musa paradisiacae pulp homogenates respectively.

Groups 5 and 6: 200 and 400mg/kg unripe Musa paradisiacae peel homogenates respectively; group 7: combination of unripe Musa paradisiacae pulp and peel homogenates (200/400 mg/kg respectively).

After the treatments, diarrhoea was  induced using castor oil. Relative to the control group 1, the treatment groups 2-7 inhibited castor oil-induced frequency of defecation and wetness of stool dose dependently but non-significantly (p>0.05).

Both the pulp and peel homogenates produced non-significant decreases (p>0.05) in the distances travelled by the charcoal meal (marker) in castor oil-induced diarrhoea rats compared to the control group 1.

Pre-treatment of the rats with unripe Musa paradisiacae pulp and peel homogenates decreased significantly (p<0.05) enteropooling indicated by decreases in the volume and weight of the gastro-intestinal contents relative to the control group 1.

Treatment with the unripe Musa paradisiacae pulp and peel homogenates led to significant decreases (p<0.05) in the bicarbonate ion concentrations except in group 3  rats while the potassium ion concentrations increased significantly (p<0.05) in all the groups except in groups 3, 4 and 7 rats which showed non-significant decreases (p>0.05) compared to the control group 1.

Sodium ion concentrations of the pre-treated groups increased non-significantly (p>0.05) except in groups 4 and 7 rats which decreased non-significantly (p>0.05) relative to the control group 1.

Using everted rat intestines, the pulp and peel homogenates enhanced significant (p<0.05) influx of sodium ions  into the everted sacs (serosal) and significant (p<0.05) efflux of potassium ions out of the sacs (mucosal) in relation to the control group 1.

These findings reveal that unripe Musa paradisiacae pulp and peel exhibit antidiarrhoeal properties by inhibiting gastro-intestinal motility, enteropooling, wetness and frequency of defecation. They have also shown abilities to facilitate transport of electrolytes across the  small  intestinal membrane.

TABLE OF CONTENTS

Title Page i
Certification ii
Dedication iii
Acknowledgements iv
Abstract v
Table of Contents vi
List of Figures xii
List of Tables xiv
List of Abbreviations xv

CHAPTER ONE: INTRODUCTION

1.1 Musa paradisiacae – – – – – – -3
1.1.1 Taxonomy of Musa paradisiacae – – – – – -5
1.1.2 Common names of Musa paradisiacae – – – – -5
1.1.3 Origin of Musa paradisiacae – – – – – – -5
1.1.4 Description of Musa paradisiacae plant – – – – -6
1.1.5 Distribution of Musa paradisiacae – – – – – -7
1.1.6 Cultivation and storage of Musa paradisiacae – – -8
1.1.7 Historical Uses of Musa paradisiacae – – – – -8
1.1.8 Health benefits of Musa Paradisiacae – – – – -8
1.1.9 Ripening process and the chemical composition of Musa paradisiacae -9
1.2 Diarrhoea – – – – – – – – -11
1.2.1 Definition of diarrhoea – – – – – – -11
1.2.2 Mechanism/pathology of diarrhoea – – – – – -12
1.2.3 Classification of diarrhoea disease – – – – – -12
1.2.3.1 Classification based on mode of infection – – – – -12
1.2.3.2 Classification based on duration of symptoms – – – -12
1.2.3.3 Classification based on pathological mechanisms – – – -13
1.2.4 Pathophysiology of diarrhoea – – – – – -13
1.2.4.1 Intestinal inflammation and diarrhoea – – – – -13
1.2.4.2 Oxidative damage in diarrhoea — – – – -15
1.2.4.3 Enteric nervous system in diarrhoea – – – – -16
1.2.4.4 Cystic fibrosis trans-membrane conductance regulator (CFTR) regulation -17
1.2.5 Specific agents of diarrhoea – – – – – – -17
1.2.6 Treatment of diarrhoea – – – – – – -19
1.2.7 Potential mechanisms in the control of diarrhoea – – – -20
1.2.8 Physiological basis of diarrhoea – – – – -23
1.3 Phytochemicals – – – – – – – -23
1.3.1 Terpenoids – – – – – – – – -24
1.3.2 Alkaloids – – – – – – – – -25
1.3.3 Phenolics – – – – – – – – -25
1.3.4 Glycosides – – – – – – – – -26
1.3.5 Tannins – – – – – – – – -26
1.4 The nervous system – – – – – – – -27
1.5 Neurotransmitters and pharmacology- – – – – -29
1.6 Some biologically important electrolytes; sodium, potassium and bicarbonates 30
1.6.1 Hyponatremia – – – – – – – – -30
1.6.1.1 Signs and symptoms of hyponatremia – – – – -30
1.6.1.2 Clinical diagnosis of hyponatremia – – – – – -31
1.6.1.3 Causes of hypernatremia – – – – – – -31
1.6.2 Hypernatremia – – – – – – – -31
1.6.2.1 Signs and symptoms of hypernatremia – – – – -31
1.6.2.2 Clinical diagnosis of hypernatremia – – – – -31
1.6.3 Hypokalemia – – – – – – – – -30
1.6.3.1 Signs and symptoms of hypokalemia – – – – -32
1.6.3.2 Clinical diagnosis of hypokalemia – – – – -32
1.6.4 Hyperkalemia – – – – – – – – -32
1.6.4.1 Signs and symptoms of hyperkalemia – – – – -32
1.6.4.2 Clinical diagnosis of hyperkalemia – – – – -33
1.7 Normal physiology of gut fluid and electrolyte transport – – -31
1.8 Electrolyte transport in the jejunum and ileum – – – -34
1.9 Castor oil – – – – – – – -35
1.9.1 Ricin – – – – – – – – – -34
1.9.2 Uses of castor oil – – – – – – -36
1.10 Antidiarrhoeal drugs – – – – – – -37
1.10.1 Nitric oxide synthase inhibitors – – – – – -37
1.10.2 Nufenoxole – – – – – – – – -37
1.10.3 Loperamide – – – – – – – – -37
1.10.4 Diphenoxylate – – – – – – – – -38
1.11 Aim of the study – – – – – – – – -38
1.12 Specific objectives of the study – – – – – – -39

CHAPTER TWO: MATERIALS AND METHODS

2.1 Materials – – – – – – – – -40
2.1.2 Experimental animals – – – – – – -40
2.1.3 Equipment – – – – – – – – -40
2.1.4 Chemical and reagents – – – – – – -40
2.2 Methods – – – – – – – – -41
2.2.1 Plant collection and identification – – – – – -41
2.2.2 Preparation of plant materials – – – – – – -41
2.2.3 Preparation of reagents – – – – – – – -41
2.2.3.1 Charcoal meal – – – – – – – – -41
2.2.3.2 5% (w/v) ferric chloride solution – – – – – -41
2.2.3.3 Ammonium solution – – – – – – – -41
2.2.3.4 45% (v/v) ethanol – – – – – – – -41
2.2.3.5 Aluminium chloride solution – – – – – – -41
2.2.3.6 Dilute sulphuric acid – – – – – – – -41
2.2.3.7 Lead acetate solution – – – – – – – -42
2.2.3.8 Wagner’s reagent – – – – – – – -42
2.2.3.9 Mayer’s reagent – – – – – – – -42
2.2.3.10 Dragendorff’s reagent – – – – – – -42
2.2.3.11 2% (v/v) hydrochloric acid – – – – – – -42
2.2.3.12 Normal saline – – – – – – – -42
2.2.3.13 1% (w/v) Picric acid – – – – – – – -42
2.2.3.14 Krebs buffer – – – – – – – – -42
2.2.3.15 0.3% glucose saline – – – – – – – -43
2.2.3.16 Preparation of standards – – – – – – -43
2.2.3.16.1 Potassium standard- – – – – – – -43
2.2.3.16.2 Sodium standard – – – – – – – -43
2.2.4 Qualitative phytochemical analysis of unripe Musa paradisiacae pulp and peel 43
2.2.4.1 Test for alkaloids – – – – – – – -43
2.2.4.2 Test for flavonoids – – – – – – – -43
2.2.4.3 Test for glycosides – – – – – – – -44
2.2.4.4 Test for saponins – – – – – – – -44
2.2.4.5 Test for tannins – – – – – – – -44
2.2.4.6 Test for terpenoids and steroids – – – – – -45
2.2.4.7 Test for reducing sugars – – – – – – -45
2.2.4.8 Test for carbohydrate – – – – – – – -45
2.2.5 Quantitative phytochemical analysis of unripe Musa paradisiacae pulp and peel 45
2.2.5.1 Quantitative determination of alkaloids – – – – -45
2.2.5.2 Quantitative determination of flavonoids – – – – -46
2.2.5.3 Quantitative determination of steroids – – – – -46
2.2.5.4 Quantitative determination of cyanogenic glycosides – – -46
2.2.5.5 Quantitative determination of tannins- – – – – -46
2.2.6 Proximate analysis – – – – – – – -47
2.2.6 1 Crude protein determination – – – – – – -47
2.2.6.2 Moisture content – – – – – – – -48
2.2.6.3 Ash – – – – – – – – – -48
2.2.6.4 Crude fibre – – – – – – – – -49
2.2.6.5 Crude fat – – – – – – – – -49
2.2.7 Determination of sodium and potassium concentration- – – -50
2.2.8 Determination of glucose – – – – – – -50
2.2.9 Acute toxicity test of aqueous extracts of unripe Musa paradisiacae pulp and peel 51
2.2.10 Anti-diarrhoeal studies – – – – – – -51
2.2.10.1 Castor oil-induced diarrhoea test – – – – – -51
2.2.10.2 Castor oil-induced enteropooling test – – – – -52
2.2.10.3 Gastro intestinal motility test – – – – – -53
2.2.10.4 Electrolyte tests – – – – – – – 53
2.2.10.4.1 Determination of sodium ion (Na+) concentration (Teco diagnostic kit) 54
2.2.10.4.2 Determination of potassium ion (K+) concentration (Teco diagnostic kit) 55
2.2.10.4.3 Determination of bicarbonate (HCO3-) ion concentration – – -55
2.2.10.5 Determination of the effects of the homogenates on the transport of sodium and potassium ions across the everted rat intestine – -56
2.2.10.5.1 Preparation of tissue – – – – – – -56
2.2.10.5.2 Preparation of the everted sacs – – – – – -56
2.2.10.5.3 Filling of the everted sacs – – – – – – -57
2.3 Statistical analysis – – – – – – – -58

CHAPTER THREE: RESULTS

3.1 The median lethal dose of unripe Musa paradisiacae pulp and peel homogenates – -59
3.2 Qualitative phytochemical composition of the unripe Musa paradisiacae pulp and peel homogenates 61
3.3 Quantitative phytochemical composition of unripe Musa paradisiacae pulp and peel homogenates-63
3.4 Proximate composition of unripe Musa paradisiacae pulp and peel – – 65
3.5 Effects of unripe Musa paradisiacae pulp and peel homogenates on castor oil-induced diarrhoea 67
3.6 Effects of unripe Musa paradisiacae pulp and peel homogenates on enteropooling in Wistar albino rats 69
3.6.1 Volume of the intestinal content of Wistar albino rats– – – -69
3.6.2 Weight of the intestinal content of Wistar albino rats- – – – -71
3.7 Effects of unripe Musa paradisiacae pulp and peel homogenates on gastro intestinal motility in Wistar albino rats – – – – 73
3.8 Effects of unripe Musa paradisiacae pulp and peel homogenates on the bicarbonate levels of the small intestinal contents of Wistar albino rats – 75
3.9 Effects of unripe Musa paradisiacae pulp and peel homogenates on the potassium ion concentrations of the small intestinal contents of Wistar albino rats – – – – – – – – 77
3.10 Effects of the of unripe Musa paradisiacae pulp and peel Homogenates on the sodium concentrations of the small intestinal contents of Wistar albino rats – – – – -79
3.11 The effects of unripe Musa paradisiacae pulp and peel homogenates on the transport of sodium ions across the everted small intestinal sac – -81
3.12 The effects of unripe Musa paradisiacae pulp and peel homogenates on the transport of potassium ion across the everted small intestinal sac – -83

CHAPTER FOUR: DISCUSSION

4.1 Discussion – – – – – – – – -85
4.2 Conclusion – – – – – – – – -88
4.3 Suggestions for Further Studies – – – – – -89
REFERENCES – – – – – – – – -90

INTRODUCTION

The use of traditional medicines in West Africa is probably as old as the duration of human settlement in the region (Abdul-aguye, 1997). A medicinal plant provides an important source of new chemical substances with potential therapeutic effects.

These have been used in traditional medicine for the treatment of several diseases and aliments (Mukerjee et al., 1998). It is already important to the global economy with demand steadily increasing not only in developing countries but also in industrialized countries (Sofowara, 1993).

Herbalism or herbal medicine is the use of plants for  medicinal purposes, and the study of such use (Briskin, 2000). Herbal medicine  is  still the  mainstay of  about 75 – 80% of the world population, mainly in the developing countries, for primary health care (Kamboj, 2000).

Plants have been the basis for medical treatments through much of human history, and such traditional medicine is still widely practiced today (Briskin, 2000).

This is primarily because of the general belief that herbal drugs are without any side effects besides being cheap and locally available (Gupta and Raina, 1998).

Modern medicine recognizes herbalism as a  form  of  alternative medicine as  the practice of herbalism is not strictly based on evidence gathered using the scientific method (Talalay, 2001).

According to  the  World  Health Organization  (WHO),  the use of herbal remedies throughout the world exceeds that of the conventional drugs by two to three times (Evans, 1994).

The use of plants for healing purposes  predates human history and forms the origin of much modern medicine.

REFERENCES 

Abdul-aguye, I. (1997). Medicinal herbs in West Africa. Annual Regional Conference of West Africa Society of Pharmacognosy, Usman Danfodio  University, Sokoto, Nigeria 22-25.

Adegboyega, O.K. (2006). Chemical composition of unripe (green) and ripe plantain (Musa paradisiaca). Journal of Science, Food and Agriculture, 24(6): 703- 707.

Adeniyi, T.A., Sanni, L.O., Barimalaa, L.S. and Hart, A.D. (2006). Determination of micronutrients and colour variability among new plantain and banana hybrid flour. World Journal of Chemistry, 1(1): 23-27.

Adesina, S.K. (1982). Studies on Nigerian herbal anti-convulsant recipe. International Journal of Crude Research, 20: 431-434.

Adeyemi, O.O. and Akindele, A.J. (2008). Antidiarrhoeal activity of the  ethyl acetate of Baphia nitida (Papilionaceae). Journal of Ethanopharmacology. 116: 407 – 412.

Agarwal, R., Afzalpurkar, R. and Fordtran, J.S. (1994). Pathophysiology of potassium absorption and secretion by the human intestine. Gastroenterology. 107: 548- 549.