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 14th 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 14th 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 15th 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
4.1.14 Histopathology
of the liver of the Wistar albino rat that received high dose of extract only 45
4.1.15 Histopathology
of the liver of the Wistar albino rat that received low dose of extract and
MSG 46
4.1.16 Histopathology
of the liver of the Wistar albino rat that received high dose of extract and
MSG 47
4.1.17 Histopathology
of the liver of the Wistar albino rat that received MSG and low dose of extract
48
4.1.18 Histopathology
of the liver of the Wistar albino rat that received MSG and high dose of
extract 49
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
LIST OF TABLE
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
Plate 4: Histopathology of the liver of
the Wistar albino rat that received low dose of extract only 44
Plate 5: Histopathology of the liver of
the Wistar albino rat that received high dose of extract only 45
Plate 6: Histopathology of the liver of
the Wistar albino rat that received low dose of extract and MSG 46
Plate 7: Histopathology of the liver of
the Wistar albino rat that receivedhigh dose of extract and MSG 47
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
CHAPTER
ONE
INTRODUCTION
Since the beginning of human
civilization, nature has provided many things for human including the tools for
the first attempt at the therapeutic intervention. During the last century, the
practice of herbalism became the popular throughout the world. In spite of the
great advances achieved in contemporary medicine, plants still make a
significance contribution to health care (Janakiramanet al., 2012). With the development of pharmaceutical industries,
much more interest has been created on plant products. They have attempted to
isolated active constituents from different plant parts and use them directly
as drugs or design them pharmacologically active compounds with or without
addition of synthetic ones (Oyiakoet al.,
2012).
Phytochemical
compounds found in plants are not required for normal functioning of the body,
but have beneficial effects on health and plays an active role in amelioration
of diseases. This is due to increased awareness of the limited ability of the
synthetic pharmaceutical products to control major diseases and the need to
discover new molecular and active structures for synthetic fields are provided
by rich natural sources (Rajibet al.,
2009).
The
family Acanthaceae consists of a significant number of medicinal plants with
broad array of biological activities and attractive number of
phytoconstituents. Asystasia, member
of the Acanthaceae family, is a genus comprising of about 70 subtropical
regions. Asystasiagangetica(L.)T.
Anderson is a fast growing, spreading, perennial herb that grows from 300-600
mm in height. It has green, oval-shaped leaves with white –cream coloured
flower with purple markings and the fruit is a club shaped capsule, splitting
from tip to base. It is native to tropical Africa, Arabic and tropical Asia,
but has been introduced in many other region where it has often naturalized. It
is widely distributed throughout the world. The plant is used in ethno medicine
for the treatment of heart pains, stomach pains, asthma, rheumatism e.t.c (Aka et al., 2003).
In Nigeria, the leaves are popularly used in
the treatment of asthma. In traditional medicine of East Africa, Asystasiagangetica is used as an
anthelminthic. Pharmacological studies have shown that the leaves ofAsystasiagangetica possess
bronchospamolytic and anti-inflammatory properties. The leaf extracts inhibited
histamine and serotonin-induced contractions of the guinea pig trachea
(Janakiramanet al., 2012). The leaves
have been shown to contain large amounts of proteins, amino acids, minerals,
carbohydrate, lipids and fibre.
1.1 Aim of study
This
study was aimed at investigating liver function indices of monosodium glutamate
induced male Wistaralbino rats treated with methanol extract of Asystasiagangetica leaves.
1.1.1 Specific objective of study
This
study was designed to achieve the following specific objectives:
i.
To determine acute toxicity of methanol
extract of Asystasiagangeticaleaves
ii.
To determine the effects of methanol
extract of Asystasiagangetica leaves
on liver marker enzymes (AST, ALT and ALP) of monosodium glutamate induced male
Wistar albino rat.
iii.
To determine the effects of the extract
on total protein, albumin, total bilirubin and conjugated bilirubin
concentration of monosodium glutamate induce male Wistar albino rats.
1.2 Problem statement
Monosodium
glutamate (MSG) commonly known with the brand name “Ajinomoto” is used for
flavour enhancer in a variety of food prepared both at homes, restaurants and
by food processors in most parts of the world including Nigeria. Its toxic
effects such as diabetes, liver damage, high blood pressure, excessive weight
gain and other health problems have been shown in numerous animal studies.
1.3 Justification of the study
Many
research works have been done on screening of some medicinal plants with
hepatoprotective and curative properties (Hewawassanet al., 2004;Buraimohet al., 2010;Adewusiet al., 2010; Akramet al., 2012;Srinivasaet al.,2012).
However, no scientifically proven information is available on the liver
function indices of monosodium glutamate induced male Wistar albino rats
treated with methanol extract of Asystasiagangeticaleaves.
Furthermore, natural products still represent an important source of
interesting leads for drugs development. While the cost of orthodox medicine
remains high and not easily affordable by the poor masses, phytomedicine which
are otherwise cheap and easily affordable, remains the first point of call for
poor patients suffering from effects associated with monosodium glutamate
consumption. Therefore, the search and discovery of new novel plants with
hepatoprotective and curative properties will likely bring hope to many people
afflicted with diseases originating from monosodium glutamate usage such as
liver damage etc.
1.4 List of abbreviation
MSG:
Monosodium
Glutamate
m/L: Microliter
ML: Millilitre
Mm: Milimetre
Kg: Kilogram
Mg: Miligram
g: gram
ALT: Alanine aminotransferase
AST:
Aspartate
amino transferase
ALP: Alkaline phosphatase
LD50: Lethal dose
B/W:
Body
weight
Fig: Figure
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