ABSTRACT

The objective of this study is to evaluate the liver function indices of monosodium glutamate induced male Wistar albino rats treated with methanol extracts of Asystasia gangetica. The acute toxicity study was carried out using 18 male Wistar albino mice while 36 male Wistar albino rats were used for the effects of the extract on liver function indices. The rats were grouped into 9 groups of 4 rats each. Group1 served as normal control and received 2 ml/kg b. wt. of distilled water for 14 days. Group 2 received a suspension of 8 g/kg b. wt. of MSG on day 7 and 14 and served as positive control. Group 3 received a suspension of 8 g/kg b. wt. of MSG and treated with 200 mg/kg b. wt. of sylimarin for 14 days. Group 4 and 5 received 200 and 500 mg/kg b. wt.  of A. gangetica leaves respectively for 14 days. Group 6 and 7 received 200 and 500 mg/kg b. wt. of A. gangetica leaves respectively for 7 days prior to MSG induction on the 7th and 14^th day respectively and continued receiving the extract for the next 7 days. Group 8 and 9 received a suspension 8 g/kg b. wt. of MSG on the first day and 14^th day and was treated with the 200 and 500 mg/kg b. wt. respectively of methanol extract of A. gangetica leaves. Blood samples and organs were collected on the 15^th day for the liver marker and function indices respectively and to ascertain the histo-architecture. The result from the acute toxicity study of methanol extract of A. gangetica leaves showed absence of death or adverse reaction which may be as a result of the methanol extract possessing low toxicity potential or that the methanol extract was relatively non-toxic. The study showed that administration of MSG caused a significant (p<0.05) increase in the liver marker enzymes (ALT, /AST and ALP) respectively and a significant (p<0.05) decrease liver function indices (total protein, conjugated bilirubin and albumin) respectively in group 2 untreated rats. However, dose dependent administration of methanol extract of A. gangetica leaves resulted to a significant (p<0.05) decrease in the liver marker enzymes and a significant (p<0.05) increase in liver function parameters respectively when compared to group 2 untreated rats. Histopathological examination of the liver showed severe infiltrations of mononuclear inflammatory leucocytes around the portal triads in group 2 untreated rats. On the other hand histopathology of the liver of groups that received dose dependent administration of the extract showed normal histomorphology of the liver. This could be attributed to methanol extract of A. gangetica leaves possessing sufficient antioxidant components which was able to reverse the effects of MSG induction with levels greater than that treated with the standard drug silymarin. Furthermore, this suggests that the extract could be used in management of health conditions associated liver dysfunction as this study shows that the extract possesses essential phyto-chemicals that exerts the hepatoprotective and curative effects observed in the liver function indices of monosodium glutamate induced male Wistar albino rats treated with methanol of Asystasia gangetica leaves.

TABLE OF CONTENT

Title page                                                                                                                                i

Declaration                                                                                                                              ii

Certification                                                                                                                            iii

Dedication                                                                                                                              iv

Acknowledgements                                                                                                                v

Table of contents                                                                                                                    vi

List of table                                                                                                                             x

List of figures                                                                                                                         xi

List of plates                                                                                                                           xii

Abstract                                                                                                                                  xiii

CHAPTER ONE:INTRODUCTION                                                                                 1

1.1 Aim of study                                                                                                                     2

1.1.1 Specific objective of study                                                                                            2

1.2       Problem statement                                                                                                      2

1.3       Justification of the study                                                                                            2

1.4.1        List of abbreviation                                                                                                     3

CHAPTER TWO: LITERATURE REVIEW

2.1       Asystasia gangetica                                                                                                     4

2.1.1    Leaves of Asystasia gangetica.                                                                                   4

2.1.2    Scientific classification of Asystasia gangetica                                                          5

2.1.3.   Botanical description of Asystasia gangetica                                                             5

2.1.4    Morphological description of Asystasia gangetica                                                     5

2.1.5.   Geographical description of Asystasia gangetica                                                       6

2.1.6    General uses of Asystasia gangetica                                                                           6

2.1.7    Traditional uses of Asystasia gangetica.                                                                     6

2.1.8    Medical uses of Asystasia gangetica.                                                                          7

2.2       Phytochemistry                                                                                                           7

2.2.1    Phytochemical Constituents                                                                                       8

2.3.1        Liver and its biological functions                                                                               8

2.3.1    Liver function parameters                                                                                           9

 2.3.1.1            Alanine amino transferase (ALT)                                                                   10

2.3.1.2             Aspartate amino transferase (AST)                                                                 10

2.3.1.3             Alkaline phosphatase (ALP)                                                                           10

2.3.1.4             Total protein                                                                                                    11

2.3.1.5             Albumin                                                                                                          11

2.3.1.6             Total bilirubin                                                                                                  12

2.3.1.7              Conjugated bilirubin                                                                                       12

2.4                                      Histopathology of liver                                                                                   12

2.5                                      Hepatotoxicity                                                                                                            13

2.5.1                Hepatoprotection                                                                                            13

2.5.2                Hepatocurative                                                                                                14

2.6                   Monosodium glutamate                                                                                  14

2.6.1                Metabolism of monosodium glutamate                                                          14

2.6.2                Hepatotoxic effects of monosodium on liver                                                 15

 2.7                  Acute toxicity                                                                                                 15

CHAPTER THREE: MATERIALS AND METHOD

3.1                   Materials                                                                                                         17

3.1.1                Plant material                                                                                                  17

3.1.2                Experimental animals                                                                                      17

3.1.3                Equipment/ apparatus                                                                                     17

3.1.4                Chemicals and reagents                                                                                  17

3.2                   Methods                                                                                                          18

3.2.1                Collection and identification of plant material                                               18

3.2.2                Collection of experimental animal                                                                  18

3.2.3                Preparation of plant material                                                                           18

3.2.4                Extraction of plant materials                                                                           18

3.2.5                Acclimatization of experimental animals                                                        19

3.2.6                Experimental design                                                                                       19

3.2.7                Preparation of solution and reagents                                                              20

3.2.7.1             Preparation of normal saline                                                                          20

3.2.7.3             Preparation of ALP reagent                                                                            20

3.2.7.4             Preparation of biuret reagent                                                                          20

3.2.7.5             Preparation of 2,4 Dinitrophenylhydrazine                                                    20

3.2.7.6             Preparation of phosphate buffer (pH7.4)                                                        20

3.2.7.7             Preparation of L- alanine substrate                                                                 20

3.2.7.8             Preparation of aspartate substrate                                                                   21

3.2.7.9             Preparation of stock biuret solution                                                                21

3.2.8                Biochemical analysis                                                                                       21

3.2.8.1             Determination of acute toxicity                                                                      21

3.2.8.2             Determination of serum ALT activity                                                            21

3.2.8.3             Determination of serum AST activity                                                             22

3.2.8.4             Determination of serum ALPactivity                                                             22

3.2.8.5             Determination of serumtotal protein concentration                                        23

3.2.8.6             Determination of total and conjugated bilirubin                                            23

3.2.8.7             Determination of serum albumin                                                                    24

3.2.8.8             Histopathological analysis of the liver                                                            24

3.3                   Statistical analysis                                                                                           25

CHAPTER FOUR: RESULTS AND DISCUSSION

4.1       Results                                                                                                                        26

4.1.1    Percentage yield                                                                                                          26

4.1.2    Lethal Dose LD₅₀toxicity of methanol extract of Asystasia gangetica Leaves          26

4.1.3    AST activity of MSG induced male Wistar albino rats                                              27

4.1.4    Effect of methanol extract ofAsystasia gangetica on ALT activity                           29

4.1.5    Effect of methanol extract of Asystasia gangetica on ALP activity                          31

4.1.6    Effect of methanol extract of Asystasia gangeticaon total protein    concentration  33

4.1.7    Effect of methanol extract of Asystasia gangetica on albumin                                  35

4.1.8    Effect methanol extract ofAsystasia gangetica total bilirubin                                    37

4.1.9    Effect methanol extract of Asystasia gangetica direct bilirubin                                 39

4.1.10  Histopathology of the liver of Wistar albino rat (control)                                          41

4.1.11    Histopathology of the liver of the Wistar albino rat that received MSG only         42

4.1.12 Histopathology of the liver of the Wistar albino rat that was induced with MSG and treated with Silymarin                                                                                                             43

4.1.13 Histopathology of the liver of the Wistar albino rat that received low dose of extract only                                                                                                                                     44                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            47

4.1.14 Histopathology of the liver of the Wistar albino rat that received high dose of extract only                                                                                                                                     45                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            48

4.1.15 Histopathology of the liver of the Wistar albino rat that received low dose of extract and MSG                                                                                                                                      46                                                                                                                                                                                                                                                                                                                                                                                                                                                        49

4.1.16 Histopathology of the liver of the Wistar albino rat that received high dose of extract and MSG                                                                                                                                      47                                                                                                                                                                                                                                                                                                                                                                                                                                                        50

4.1.17 Histopathology of the liver of the Wistar albino rat that received MSG and low dose of extract                                                                                                                                     48                                                                                                                                                                                                                                                                                                                                                                                                                                                        51

4.1.18  Histopathology of the liver of the Wistar albino rat that received MSG and high dose of extract                                                                                                                                     49                                                                                                                                                                                                                                                                                                                                                                                                                                                        52

4.2       Discussion                                                                                                                   50

CHAPTER FIVE: CONCLUSION AND RECOMMENDATION

5.1       Conclusion                                                                                                                  55

5.2       Recommendation                                                                                                        56

5.3       References                                                                                                                  52

5.4       Appendix                                                                                                                    59

Table 1: Acute toxicity of Methanol Extract of Asystasia gangetica                                     26

LIST OF FIGURES

Fig.1: Aspatate amino transferase activity of monosodium glutamate induced male Wistaralbino rats treated with methanol extract of Asystasia gangetica                                                28

Fig. 2: Alanine amino transferase activity of monosodium glutamate induced male Wistar albino rats treated methanol extract of Asystasia gangetica                                                        30

Fig. 3: Alkaline phosphatase (ALP) amino transferase activity of monosodium glutamate induced male Wistar albino rats treated with methanol extract of Asystasia gangetica                   32

Fig. 4: Total protein concentration of monosodium glutamate induced male Wistar  albino rats treated with methanol extract of Asystasia gangetica                                                            34

Fig. 5: Mean albumin concentration of monosodium glutamate induced male Wistar albino rats treated with methanol extract of Asystasia gangetica                                                            36                                                                                                                                             39

Fig. 6: Total bilirubin concentration of monosodium glutamate induced male Wistar albino rats treated with methanol extract ofAsystasia gangetica                                                             38

Fig. 7: Direct bilirubin concentration of monosodium glutamate inducedmale Wistar albino rats treated with methanol extract of Asystasia gangetica                                                            40

LIST OF PLATES

Plate 1: Histopathology of the liver of Wistar albino rat (control)                                         41

Plate 2: Histopathology of the liver of the Wistar albino rat that received MSG only          42

Plate 3: Histopathology of the liver of the Wistar albino rat that was  induced with MSG and treated with Silymarin                                                                                                             43                                                                                                                                                                                                                                                                                                                                                                                                                                                        46                                                                                                                                   

Plate 4: Histopathology of the liver of the Wistar albino rat that received low dose of extract only                                                                                                                                    44                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        47

Plate 5: Histopathology of the liver of the Wistar albino rat that received high dose of extract only                                                                                                                                     45                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        48

Plate 6: Histopathology of the liver of the Wistar albino rat that received low dose of extract and MSG                                                                                                                                     46                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            49

Plate 7: Histopathology of the liver of the Wistar albino rat that receivedhigh dose of extract and MSG                                                                                                                                     47                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            50

Plate 8: Histopathology of the liver of the Wistar albino rat that received MSG and low dose of extract                                                                                                                                     48                                                                                                                                                                                                                                                                                                                                                                                                                                                        51

Plate 9: Histopathology of the liver of the Wistar albino rat that received

 MSG and high dose of extract                                                                                  49