TABLE OF CONTENTS
Title page
Table of contents
List of Abbreviation
Abstract
CHAPTER ONE
1.0 INTRODUCTION
1.1 Background
1.2 Statement of Problem
1.3 Justification
1.4 Significance of Study
1.5 Aim and Objectives of the Study
1.5.1 Aim of the study
1.5.2 Objectives of the study
1.6 Hypothesis
CHAPTER TWO
2.0 LITERATURE REVIEW
2.1 Lawsonia inermis
2.1.1 Taxonomy of Lawsonia inermis
2.1.2 Origin and distribution of Lawsonia inermis
2.1.3 Local names of Lawsonia inermis
2.1.4 Morphology of Lawsonia inermis
2.1.5 Phytochemistry of Lawsonia inermis
2.1.6 Ethnomedicinal use of Lawsonia inermis
2.1.7 Pharmacology of Lawsonia inermis
2.1.8 Safety evaluation of Lawsonia inermis
2.1.9 Ethnobotanical uses of Lawsonia inermis
2.2 The Liver
2.2.1 Embryology of the liver
2.2.2 Gross anatomy of the liver
2.2.3 Histology of the liver
2.2.4 Functions of the liver
2.2.5 Liver diseases
2.3 Ethanol
2.3.1 Uses of ethanol
2.3.2 Pharmacology of ethanol
CHAPTER THREE
3.0 MATERIALS AND METHOD
3.1 Materials
3.1.1 Collection and identification of plant
3.1.2 Experimental animals
3.1.3 Equipment/instruments
3.1.4 Reagents
3.2 Methods
3.2.1 Preparation of plant extract
3.2.2 Determination of dose
3.2.3 Experimental design
3.2.4 Hepatotoxicity induction
3.2.5 Sacrifice of animals
3.2.6 Morphological studies
3.2.7 Biochemical studies
3.2.8 Molecular study
3.2.9 Histological studies
3.2.10 Statistical analysis
CHAPTER FOUR
4.0 RESULTS
4.1 General Observation
4.2 Morphological Studies
4.2.1 Body weight change
4.2.2 Liver-body weight ratio
4.3 Biochemical Studies
4.3.1 Oxidative stress markers
4.3.2 Protein concentration
4.4 Molecular Study
4.4.1 DNA fragmentation
4.5 Histological Studies
4.5.1 Haematoxylin and eosin (H&E) staining
4.5.2 Acridine orange and ethidium bromide (AO-EB) staining
CHAPTER FIVE
5.0 DISCUSSION
5.1 Morphological Studies
5.2 Biochemical Studies
5.2.1 Oxidative stress markers
5.2.2 Protein concentration
5.3 Molecular Study (DNA Fragmentation)
5.4 Histological Study
CHAPTER SIX
6.0 CONCLUSION AND RECOMMENDATIONS
6.1 Conclusions
6.2 Recommendations
CONTRIBUTIONS TO KNOWLEDGE
REFERENCES
ABSTRACT
Lawsonia inermis commonly known as henna has been used traditionally,
especially in ayurvedic medicine, for various conditions including liver
ailments, and reported to have hepatoprotective properties. This study aims to
study the effect of aqueous extract of L. inermis leaves in acute
ethanol induced hepatic damage in adult Wistar rats. Thirty (30) female
rats were equally divided into five (5) groups (I-V). Group I which served as
negative control received distilled water (2 ml) for 8 days. Group II which
served as positive control received 40 % ethanol (20 ml/kg) on the 8th
day after seven days of distilled water. Group III which served as prophylactic
group received 400 mg/kg of aqueous extract of Lawsonia inermis for
seven days, and 40 % ethanol (20 ml/kg) on the 8th.
Group IV served as the therapeutic group, and received 400 mg/kg of the extract
for seven days after receiving 20 ml/kg of 40 % ethanol on the first. Group V
received silymarin (70 mg/kg) for seven days before 20 ml/kg of 40 % ethanol on
the 8th
day, to serve as the reference drug. All animals were sacrificed on the ninth
(9) day. Body weight changes and liver body weight index were determined. Liver
tissues were collected for assessment of oxidative stress markers (superoxide
dismutase, catalase, reduced glutathione and malondialdehyde), protein
concentration, DNA fragmentation, and also for haematoxylin and eosin staining,
and acridine orange-ethidium bromide staining. Body weight increased in all
groups from initial mean weight of 121.5 g, though significantly (p<0.05)
only in Group I and Group IV. Liver body weight index was highest in Group II,
and was significantly (p<0.05) different from Group I and Group IV. Ethanol
administration reduced levels of Superoxide dismutase, catalase, reduced
glutathione and caused an increase in lipid peroxidation (Malondialdehyde),
though insignificantly. Increase in levels of Superoxide dismutase, catalase,
reduced glutathione and decrease in lipid peroxidation (Malondialdehyde) was
observed in the groups receiving extract, more so prophylactically,
though statistically insignificant. Silymarin significantly increased the level
of Superoxide dismutase in comparison to Group II, levels of catalase, reduced
glutathione, and Malondialdehyde were insignificantly less than Groups III and
IV. Protein concentration was significantly higher in Group II, Group III and
Group V, with Group II having the highest. Group III and Group V also differed
significantly (p<0.05) from Group II. DNA fragmentation was observed to be
most in Group II, while Group III and Group V had Fragmentation pattern
comparable to that of Group I. H&E staining revealed attenuation of the
effects of ethanol administration by the extract and silymarin, though the
extract proved more effective prophylactically at 400 mg/kg dose and duration
of seven days. Acridine orange-ethidium bromide (AO-EB) staining revealed
reduced necrotic and apoptotic cells in Groups III, IV and V. The extract of Lawsonia
inermis at 400 mg/kg for seven days proved to have more effective hepatoprotection
prophylactically than therapeutically, and provided comparable hepatoprotection
with that of silymarin.
CHAPTER ONE
1.0 INTRODUCTION
1.1 Background
Alcohol consumption is customary in most cultures and alcohol abuse is common worldwide. For example, more than 50 % of Americans consume alcohol and 23 % of Americans participated in heavy and/or binge drinking at least once in a month (Substance Abuse and Mental Health Services Administration, 2011). The most commonly recognized symptoms of alcohol consumption are associated with chronic alcoholism, and it is a casual/risk factor in about 60 major types of diseases (Massey and Arteel, 2012). These, and other effects of alcohol consumption, has made alcohol the third leading risk factor globally for disease and disability (WHO, 2011).
Acute Alcohol exposure or binge drinking is defined in National Survey on Drug Use and Health (NSDUH) 2010 as five or more drinks on a single occasion within a time period of three hours (Massey and Arteel, 2012). However, it can also include a period of heavy drinking that may span several days or periods of intermittent, repeated episodes of heavy drinking (Massey and Arteel, 2012). Reports by NSDUH show that more than 58 million Americans participated in binge drinking on at least one occasion within thirty days in 2010 (Substance Abuse and Mental Health Services Administration, 2011) and acute alcohol consumption is a major underlying cause of morbidity and mortality during hospital admittance (Jones et al., 1991). Due to this, and increasing prevalence of binge drinking in people aged 18-25 (Massey and Arteel, 2012), greater interest has turned towards the effect of acute alcohol exposure.....
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