COMPARATIVE STUDY OF THE EFFECTS OF AQUEOUS EXTRACTS OF MORINGA OLEIFERA AND VERNONIA AMYGDALINA LEAVES ON SOME BIOCHEMICAL INDICES IN ALBINO

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ABSTRACT

A comparative study of the effects of aqueous extract of Moringa oleifera and Vernonia amygdalina on selected biochemical indices of prostate pathologies of male albino rats was investigated using thirty two adult male albino rats of average weight of 152g. Rats were randomly assigned into four groups of eight rats each with different recognizable codes. Group one served as the control and each rat in this group received orally 2ml of distill water daily. Groups two and three received oral administrations of 2ml aqueous extract containing 5g of M. oleifera, V. amygdalina respectively daily. Group four received 4ml containing 2.5g of M. oleifera with 2.5g of V. amygdalina per body weight orally daily. Administration of the extract was done for 28 days. The rats were fed on growers match and tap water ad libitum throughout the experiment. The animals were housed in standard animal cages in the animal house of the Department of Biochemistry, Kogi State University, Anyigba under 12 hour light/ dark cycle for 29 days. Changes in weight were noted at the end of every week for the four weeks. The animals were starved over-night and blood drawn from cut at the tip of the tails was used for fasting blood glucose test. All the animals were killed on the 29th day by jugular puncture; part of their blood was collected into plain bottle and spun to obtain serum, and part into EDTA bottle for biochemical analyses. A non significant weight increase was observed till the third week of administration of the extract followed by a decrease in weight in the fourth week. The group administered a combination of the extracts of M. oleifera with V. amygdalina showed a significant decrease (P<0.05) when compared to the control. A significant reduction (p<0.01) in the fasting blood glucose (FBS) was recorded in the group administered M. oleifera alone and the combination of M. oleifera with V. amygdalina when compared to the control value. An increase in the Packed Cell Volume was recorded in all the groups but were not significant (P>0.05). A significant increase in the total protein was recorded for the group administered V. amygdalina and the combination of M. oleifera with V. amygdalina. A significant increase (p<0.05) and (p<0.01) in the serum globulin concentration was observed in the group administered only V. amygdalina and the group administered the combination respectively. M. oleifera increased serum albumin compared to the V. amygdalina and the combination, but decreased the serum bilirubin concentration relative to V. amygdalina and the combination. A significant increase (p<0.001) and (p<0.05) in the urea level was observed in the group administered the combination of M. oleifera and V. amygdalina and the group administered only M. oleifera respectively when compared to the control value. For serum electrolyte, there was an equivalent increase in serum potassium ion in all the groups but not significant (P>0.05) when compared to the control value. Both plant leaf extracts reduced the serum chloride ion but the combination did it better. M. oleifera recorded a significant reduction in serum sodium ion compared to V. amygdalina and the combination but their differences was not significant (P>0.05). For serum lipid profile, V. amygdalina has a significant reduction (p<0.05) potential over M. oleifera for total cholesterol. V. amygdalina and the combination increased HDL compared to M. oleifera but not significant (P>0.05). LDL decreased significantly (P<0.05) in the combined extract relative to the single. Significant decreases (p<0.001) in Prostatic Acid Phosphatase (PAP) and Total Acid Phosphatase (TAP) activities were recorded in the entire intervention group when compared to the control value. However, the two plants showed a reduction potential in metabolic factors associated with prostate disorders and their markers, PAP and TAP.

TABLE OF CONTENTS

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Title Page        ..          ..        ..          ..          ..          ..          ..          ..          i

Certification    ..          .          ..          ..          ..          ..          ..          ..          ii

Dedication      ..          ..           ..          ..          ..          ..          ..          ..          iii

Acknowledgements    ..                ..          ..          ..          ..          ..          ..          iv

Abstract          ..           ..          ..          ..          ..          ..          ..          ..          v

Table of Contents       .       ..          ..          ..          ..          ..          ..          ..          vi

List of Figures ..                    ..          ..          ..          ..          ..          ..          ..          x

List of Tables  ..           ..          ..          ..          ..          ..          ..          ..          xi

CHAPTER ONE: INTRODUCTION

1.1       Prostate          …        …        …        …        …        …        …        1

1.1.1    Prostate pathologies    …          …        …        …        …        …        …        1

1.1.1.1 Benign Prostate Hyperplasia (BPH)   …       …        …        …        …        1

1.1.2 Etiology of BPH         …        …        …        …        …        …        …        3

1.1.3 Prostatitis  …        …        …        …        …        …        …        …        4

1.1.4 Prostate cancer          …        …        …        …        …        …        …        4

1.1.5 Androgens      …        …        …        …        …        …        …        …        5

1.2       Functions of androgens         …        …        …        …        …        …        5

1.2.1 Androgens and prostate disorders             …        …        …        …        6

  1. Estrogens        …        …        …        …        …        …        …        7

1.2.3 Functions of estrogens   …              …        …        …        …        …        8

  1. Metabolic factors associated with prostate pathologies …        …        8

1.3       Prevalence of prostate pathologies            …        …        …        …        10

1.3.1 Treatment/ Management option of Prostate pathologies …        10

1.3.2 Agents that relax smooth muscles         …        …        …        …        10

1.3.3 Agents that reduce prostate volume           …        …        …        …        10

1.3.4 Medications that inhibit 5, alpha-reductase               …        …        …

1.3.5 Diet and prostate pathologies    …        …        …        …        …        …        …

1.4. Botanical/ Phytotherapy     …        …        …        …        …        …        15

1.4.1 Moringa oleifera  …        …        …        …        …        …        17

1.4.1.1 Nutrition             …        …        …        …        …        …        …        18

1.4.1.2 Phytochemistry             …        …        …        …        …        …        18

1.4.1.3 Disease treatment and prevention       …        …        …        …        18

1.4.1.4 Antibiotic activity           …        …        …        …        …        …        19

1.4.1.5 Cancer prevention      …        …        …        …        …        …        19

1.4.2 Vernonia amydalina       …        …        …        …        …        …        …        …19

1.5 Study Rationale/Justification               …        …        …        …        …20

1.6 Aim and objectives of the study          …        …        …        …        …21

1.6.1 Aim of the study        …        …        …        …        …        …        …        …21

  1. Objectives of the Study            …        …        …        …        …21

CHAPTER TWO: MATERIALS AND METHODS

2.1. Materials              …        …        …        …        …        …        …        …        …22

2.1.1 Reagents and chemicals          …        …        …        …        …        …22

2.1.2 Equipment/Devices        …      …        …        …        …        …        …22

2.1.3 Plants used         …        …        …        …        …        …        …        …        …23

2.1.4 Animals              …        …        …        …        …        …        …        …        …23

2.2 Methods                …        …        …        …        …        …        …        …        …23

2.2.1 Preparation of plant used and animal care          …        …        …23

2.2.2 Study Design     …        …        …        …        …        …        …        …        …24

2.2.3 Determination of total cholesterol concentration in serum       …24

2.2.4 Determination of high density lipoprotein (HDL) concentration in serum       …25

2.2.5 Determination of low density lipoprotein (LDL) concentration in serum         …26

2.2.6 Determination of very low density lipoprotein (VLDL) concentration in serum

2.2.7 Estimation of non-HDL           …        …        …        …        …26

2.2.8 Determination of fasting blood glucose (FBG) concentration      …27

2.2.9 Assay of total and prostatic acid phosphatases (TAP and PAP) in serum        …27

Table 4: Procedure for assaying serum total and prostatic acid phosphatise              …28

2.2.10 Determination of the concentration of total proteins in the serum      …        …28

2.2.11 Determination of the concentration of total albumin in serum            …        …29

2.2.12 Estimation of globulin in serum         …        …        …        …30

2.2.13 Packed cell volume (PCV) determination            …        …        …30

2.2.14 Determination of creatinine in serum      …        …        …        …31

2.2.15  Determination of sodium ion in serum    …        …        …        …31

2.2.16  Determination of potassium ions in serum       …        …        …32

2.2.17  Determination of chloride ions in serum         …      …        …        …33

2.2.18  Determination of urea in serum          …           …        …        …        …34

  1. Statistical analysis       …        …        …        …        …        …        …        …35

CHAPTER THREE: RESULTS

3.1       Weights of experimental rats in the study groups        …        …        36

3.2       Physical observation of the rats       …        …        …        …        39

3.3       Total protein of rats administered aqueous leaf extract of

  M. oleifera and V. amygdalina …        …        …        … 39

3.4       Serum albumin of rats administered aqueous leaf extract of

  M. oleifera and V. amygdalina  …        …        …        …        39

3.5       Serum globulin of rats administered aqueous leaf extract of

M. oleifera and V. amygdalina  …        …        …        …        39

3.6       Serum total haemoglobin of rats administered aqueous leaf extract of M. oleifera and V. amygdalina …        …        …        …        41

3.7       Packed cell volume (PCV) of rats        …        …        …        …        43

3.8       Serum bilirubin of rats of rats administered aqueous leaf extract of

M. oleifera and V. amygdalina            …        …        …        …        …        …        45

3.9       Serum creatinine of rats of rats administered aqueous leaf extract of

  M. oleifera and V. amygdalina …        …        …        …        45

3.10     Serum Urea of rats of rats administered aqueous leaf extract of

M. oleifera and V. amygdalina            …        …        …        …        …        …        45

3.11     Serum potassium ion of rats of rats administered aqueous leaf extract of M. oleifera and V. amygdalina  …        …        …        …        47

3.12     Serum chloride ion of rats of rats administered aqueous leaf extract of M. oleifera and V. amygdalina  …        …        …        …        47

3.13     Serum sodium ion of rats of rats administered aqueous leaf extract of

M. oleifera and V. amygdalina …        …        …        …        … 47

3.14     Serum lipid profile of rats of rats administered aqueous leaf extract of M. oleifera and V.  amygdalina  …        …        …        …        49

3.15     Serum total acid phosphatase (TAP) of rats of rats of rats administered aqueous leaf extract of M. oleifera and V.  amygdalina         …        …        …        51

3.16     Serum prostatic acid phosphatase (PAP) of rats of rats administered

aqueous leaf extract of M. oleifera and V. amygdalina  …        51

  • Fasting Blood Sugar (FB) of rats of rats administered aqueous leaf

extract of M. oleifera and V. amygdalina …        …        …        53

CHAPTER FOUR:  DISCUSSION

4.1       Discussion      …        …        …        …        …        …55

4.2       Suggestions for Further Research         …        …        …        …59

REFERENCES        …        …        …        …        …        …        …        …        …60   

APPENDICES          …        …        …        …        …        …        …        …        …77   

LIST OF FIGURES            

PAGE

Fig. 1: Changes in weight of rats administered M. oleifera and V. amygdalina after four weeks   …   …        …        …        …        …        …        37

Fig. 2: Percentage change in weights in rats administered M. oleifera             and V. amygdalina after four weeks  … …    …        …        …        …        …        38

Fig. 3: Serum total protein, albumin and globulin concentration of rats         administered M. oleifera and V. amygdalina after four weeks … …        40

Fig. 4: Serum total haemoglobin concentration of rats administered M. oleifera and V. amygdalina after four weeks        …        …        …        …        42

Fig. 5: Packed cell volume of rats administered M. oleifera and V. amygdalina       …        44

Fig. 6: Serum urea, bilirubin and urea concentration of rats administered M. oleifera and V. amygdalina …        …        …        …        46

Fig. 7: Serum potassium, sodium and chloride ion concentrations             of rats administered M. oleifera and V. amygdalina   …        …        …        …        48

Fig. 8: Lipid profile of rats administered M. oleifera and V. amygdalina      …        …        50

Fig. 9: Prostate markers (PAP and TAP) of rats administered             M. oleifera and V. amygdalina            …        …        …        …        …        …        52

Fig. 10: Prostate markers (PAP and TAP) of rats administered            M. oleifera and V. amygdalina            …        …        …        …        …        …        52

LIST OF TABLES

Table 1: Physical observation of rats in their various groups              …        …        …        39

                                                        CHAPTER ONE

                                                     INTRODUCTION

1.1       Prostate

The prostate is a gland found only in males. It is located in front of the rectum and below the urinary bladder and vas deferens. The prostate secretes an alkaline fluid that forms part of the semen. The size of the prostate varies with age. In younger men, it is about the size of a walnut, but it can be much larger in older men.

The prostate makes some of the fluid that protects and nourishes sperm cells in semen, making the semen more liquid. Just behind the prostate are glands called seminal vesiclesthat make most of the fluid for semen. The urethra, the tube that carries urine and semen out of the body through the penis, goes through the center of the prostate.

The prostate starts to develop before birth. It grows rapidly during puberty, fueled by male hormones (called androgens) in the body. The main androgen, testosterone, is made in the testicles. The enzyme 5-alphareductaseconverts testosterone into dihydrotestosterone(DHT). DHT is the main hormone that signals the prostate to grow (Lu-Yao, 2008).The prostate usually stays at about the same size or grows slowly in adults, as long as male hormones are present.

1.1.1    Prostate pathologies

COMPARATIVE STUDY OF THE EFFECTS OF AQUEOUS EXTRACTS OF MORINGA OLEIFERA AND VERNONIA AMYGDALINA LEAVES ON SOME BIOCHEMICAL INDICES IN ALBINO