ABSTRACT
In recent times, the cultivation of Orange Flesh Sweet Potato (OFSP) has advanced its industrial applications due to its high β-carotene and dry matter content. This study was aimed at evaluating the hepatoprotective and antihyperlipidaemic effects of OFSP juice. A total of 85 male Sprague Dawley rats (80-120g) were used for this study. Sixty rats were used for the hepatoprotective (preventive and curative) study. The animals were grouped into five groups; Group 1- Normal control, Group 2- Untreated, Group 3- Silymarin, Groups 4 – 5 (300 and 600 mg/kg body weight of OFSP, respectively). Twenty-five (25) rats were used for the antihyperlipideamic study consisting of five groups; Group 1- High fat diet control (HFDC), Group 2- Low fat diet control (LFDC), Group 3- 200 mg/kg b. w of OFSP, Group 4- 400 mg/kg b. w. of OFSP, Group 5- 600 mg/kg b. w. of OFSP. The biochemical analyses were determined using standard biochemical methods. The results indicated significant (p<0.05) decrease in the total bilirubin (TB) concentrations of the OFSP-treated preventive (12.02 ± 0.39) and curative (4.35 ± 0.21) groups compared to the untreated preventive (20.97 ± 1.27) and curative (17.22 ± 0.20) groups of the rats. There were also significant (p<0.05) decrease in the CB, AST, ALT and ALP activities in the OFSP treated groups relative to the untreated in the preventive and curative models. The haemoglobin concentration (HB) 9.51 ± 0.26, packed cell volume (PCV) 28.43 ± 0.32, red blood cell (RBC) 5.44 ± 0.25, total lymphocyte count (TLC) 11.95 ± 0.13 and platelet count (PLAT) 115.70 ± 0.85 were significantly (p<0.05) increased in the OFSP treated group when compared to the untreated and silymarin-treated group. There were no visible histological changes (alterations) in the OFSP-treated groups compared to the untreated group. For the antihyperlipidaemic study, the cholesterol concentration was 65.06 ± 10.25 in the high fat diet control, 52.24 ± 6.72 in the low fat diet control and 61.22 ± 1.52 for the OFSP treated group this showed that there was no significant (p<0.05) difference in the OFSP treated group and HFDC/LFDC. Triglyceride also showed no significant (p<0.05) difference in the OFSP (89.64±9.06) and the HFDC (105.74±16.52)/LFDC (78.50±1.67) group while High density lipoprotein (HDL) showed a significant (p<0.05) increase in the OFSP treated group (29.92±1.85) compared to the HFDC (15.56±2.10), LDL and VLDL showed a significant (0.05) decrease in the OFSP treated group compared to the HFDC. It is concluded from this study that OFSP juice can ameliorate liver damage and may have a mild antihyperlipidaemic potential.
TABLE OF CONTENTS
Title page i
Dedication ii
Acknowledgement iii
Table of Contents iv
List of Tables v
List of Figures vi
Abstract vii
CHAPTER 1
INTRODUCTION
1.1 Background of the Study 1
1.2 Statement of the Problem 3
1.3 Aim 4
1.4 Objectives 4
1.5 Justification of Study 4
CHAPTER 2
LITERATURE REVIEW
2.1 Overview of Ipomoea batatas 6
2.1.1 Scientific
classification 6
2.2 Nutritional
Constituents of Orange Flesh Sweet Potato (OFSP) 7
2.2.1 Proximate composition 7
2.2.2 Mineral constituents of OFSP 8
2.2.3 Carotenoids 11
2.2.3 Tocopherol 11
2.3 Pharmacological Properties of Ipomea batatas 12
2.3.1 Antiulcer
activity 12
2.3.2 Anti-inflammatory activity 13
2.3.3 Hepatoprotective effect 13
2.3.4 Immunomodulatory effect 13
2.3.5 Cardiovascular effect 13
2.3.6 Anti-proliferative activity 14
2.3.7 Antioxidant activity 14
2.3.8 Wound healing effect 14
2.3.9 Anti-diabetic effect 14
2.3.10 Anti-cancer
potential 15
2.3.12
Haematological effects 15
2.4 Liver Toxicity 15
2.4.1 Stages of liver toxicity/damage
16
2.4.2 Hepatotoxicity 17
2.4.3 Liver function tests
20
2.5 Mechanism of Action of Silymarin
(Hepatoprotective Agent) 23
2.6 Hepatoprotective Properties of Some
Medicinal Plants 23
2.6.1 Dodonaea
viscosa (Sapindaceae) 23
2.6.2 Phyllanthus muellarianus (Euphorbiaceae) 24
2.6.3 Aquilaria agallocha (Thymelaeaceae) 24
2.6.4 Phoenix dactylifera (Arecaceae) 25
2.6.5 Convolvulus arvensis (Convolvulaceae) 25
2.6.6 Salix caprea (Salicaceae) 26
2.6.7 Caesalpinia crista (Fabaceae) 26
2.6.8 Alocasia indica (Araceae) 26
2.6.9 Opuntia ficus-indica (Cactaceae) 27
2.6.10 Allium cepa 27
2.7 Hyperlipidemia 27
2.7.1 Risk Factors of Hyperlipidaemia 29
2.7.2 Complications of Hyperlipidaemia 30
2.8 Hypolipidaemic Properties of Some
Medicinal Plants 31
2.8.1 Lagenaria
siceraria 31
2.8.2 Cassia
angustifolia 32
2.8.3 Cinnamomum
tamala 32
2.8.4 Gymnema
sylvestre 32
2.8.5 Hibiscus
cannabinus 33
2.8.6 Glycyrriza
glabra (GG) 33
2.8.7 Moringa
oleifera 33
2.8.8 Sida
cordifolia 34
2.8.9 Spirulina
platensis 34
CHAPTER 3
MATERIALS AND METHODS
3.1 Materials 35
3.1.1 Sample procurement 35
3.1.2 Sample preparation
35
3.1.3 Chemicals/reagents 35
3.1.3 Experimental animals 35
3.2 Experimental Design 36
3.2.1 Hepatoprotective activity 36
3.2.2 Hepato-curative activity 36
3.2.3 Antihyperlipidaemic activity 37
3.3 Methods 38
3.3.1 Phytochemical screening
38
3.3.2 GC-MS analysis 40
3.3.3 Assessment of liver function 41
3.3.4 Haematology parameters
44
3.3.5 Lipid profile assays 47
3.4 Statistical Analysis 53
CHAPTER
4
RESULTS
AND DISCUSSION
4.1 Results 54
4.1.1
Qualitative phytochemical composition of OFSP juice 55
4.1.2 Chemical composition/bioactive compounds in
OFSP juice
56
4.1.3 Curative effect of OFSP on serum total bilirubin
concentration in thioacetamide-induced Liver damage in sprague dawley rat. 59
4.1.4 Curative Effect of OFSP on serum conjugate bilirubin
concentration in thioacetamide-induced liver damage in sprague dawley rat. 61
4.1.5 Curative effect of OFSP on serum aspartate
transaminaase concentration in thioacetamide-induced liver damage in sprague
dawley rat. 63
4.1.6 Curative effect of OFSP on serum alanine
transaminase concentration in thioacetamide-induced liver damage in sprague
dawley rat. 65
4.1.7 Curative effect of OFSP on serum alkaline
phosphatase concentration in thioacetamide-induced liver damage in sprague
dawley rat. 67
4.1.8 Preventive effect of OFSP on serum total
bilirubin concentration in thioacetamide-induced liver damage in sprague dawley
rat. 69
4.1.9 Preventive effect of OFSP on serum
conjugate bilirubin concentration in thioacetamide-induced liver damage in sprague
dawley rat. 71
4.1.11 Preventive effect of OFSP on serum aspartate
transaminaase concentration in thioacetamide-induced liver damage in sprague
dawley rat.
73
4.1.12 Preventive effect of OFSP on serum alanine
transaminase concentration in thioacetamide- induced liver damage in sprague
dawley rat. 75
4.1.13 Preventive effect of OFSP on serum alkaline
phosphatase concentration in thioacetamide-induced liver damage in sprague
dawley rat. 77
4.1.14 Curative effect of OFSP on some
haematological parameters 79
4.1.15 Preventive effect of OFSP on some
haematological parameters in thioacetamide-induced liver damage in sprague dawley rat 81
4.1.16 Histopathology of the liver
(hepato-curative) 83
4.1.17 Histopathology
of the Liver (Hepatoprotective) 89
4.1.18 Effect of OFSP in lipid profile parameters in
high fat diet induced hyperlipidaemia 95
4.1.19 Effect of OFSP on body weight of high fat
diet induced hyperlipidaemia sprague dawley rats
97
4.2 DISCUSSION 99
CHAPTER
5
CONCLUSION
AND RECOMMENDATION
5.1 Conclusion 108
5.2 Recommendation 108
REFERENCES
109
APPENDIX
LIST OF TABLES
3.1 Feed Formulation for Antihyperlipidaemic
Activity
38
4.1 Qualitative Phytochemical Composition of
OFSP Juice 55
4.2 Chemical Composition of Orange Flesh Sweet
Potato (OFSP)
57
4.3
Bioactive Chemical Compounds found in
Orange Flesh Potato Juice 58
4.4 Curative Effect of OFSP on Haematological
Parameters in Thioacetamide-induced Hepatotoxicity 80
4.5 Effect
of OFSP juice pretreatment on Haematological Parameters in Thioacetamide- induced Hepatotoxicity
in Sprague Dawley Rats.
82
4.6
Effect of OFSP on Lipid Profile
Parameters of High Fat Diet-induced Hyperlipidaemia 96
4.7 Effect of OFSP on Body Weight of High Fat
Diet-induced Hyperlipidaemia Sprague Dawley Rats 98
LIST
OF FIGURES
4.1
Curative Effect of OFSP on Serum Total Bilirubin Concentration in Thioacetamide-induced
Liver Damage in Sprague Dawley Rat 60
4.2 Curative Effect of OFSP on Serum
Conjugate Bilirubin Concentration in Thioacetamide-induced Liver Damage in
Sprague Dawley Rat. 62
4.3 Curative Effect of OFSP on Serum
Aspartate Transaminase Activity in Thioacetamide-induced Liver Damage in
Sprague Dawley Rat. 64
4.4 Curative
Effect of OFSP on Serum Alanine Transaminase Activity in Thioacetamide-induced Liver
Damage in Sprague Dawley Rat. 66
4.5
Curative Effect of OFSP on Serum Alkaline
Phosphatase Activity in Thioacetamide-induced Liver Damage in Wistar Albino
Rat. 68
4.6 Effect of OFSP juice pretreatment on
Serum Total Bilirubin concentration in Thioacetamide- induced hepatotoxicity in Sprague Dawley rats. 70
4.7 Effect of OFSP juice pretreatment on
Serum Conjugate Bilirubin concentration in Thioacetamide-induced
hepatotoxicity in Sprague
Dawley rats. 72
4.8 Effect of OFSP juice pretreatment on
Serum Aspartate Transaminase Activity in Thioacetamide-induced
Hepatotoxicity in Sprague
Dawley rats. 74
4.9 Effect
of OFSP juice pretreatment on Serum Alanine Transaminase Activity
Thioacetamide- induced
Hepatotoxicity in Sprague
Dawley rats. 76
4.10 Effect
of OFSP juice pretreatment on Serum Alkaline Phosphatase Activity
Thioacetamide- induced
Hepatotoxicity in Sprague
Dawley rats.
78
4.11 Histological sections of the liver of rats
given distilled water (normal rats). 84
4.12 Histological sections of the liver of rats
given thioacetamide (400 mg/kg). 85
4.13 Histological sections of the liver of rats
given thioacetamide (400 mg/kg) and 50 mg/kg silymarin. 86
4.14 Histological sections of the liver of rats
given thioacetamide (400 mg/kg) and 300 mg/kg b. w of OFSP juice. 87
4.15 Histological sections of the liver of rats
given thioacetamide (400 mg/kg) and 600 mg/kg b. w of OFSP juice.
88
4.16 Histological sections of the liver of rats
given distilled water in the pretreated group (normal
control). 90
4.17 Histological sections of the liver of rats
pretreated with thioacetamide (400 mg/kg). 91
4.18 Histological sections of the liver of rats
pretreated with 50 mg/kg silymarin and thioacetamidein the pretreated group (400 mg/kg).
92
4.19 Histological sections of the liver of rats
pretreated with 300 mg/kg b.w. and thioacetamide (400 mg/kg). 93
4.20 Histological sections of the liver of rats
pretreated with 600 mg/kg b.w. of OFSP and thioacetamide
(400 mg/kg).
94
CHAPTER 1
INTRODUCTION
1.1
BACKGROUND
OF THE STUDY
In
recent times, the cultivation of Orange Flesh Sweet Potato (OFSP) has advanced
its industrial applications due to its high β-carotene and dry matter content
adding to its traditional usage as feed and food (Nedunchezhiyan et al., 2012). Foods produced from OFSP
provide sufficient amounts of energy and β-carotene to children, pregnant and
lactating women reducing vitamin A deficiency and under-nutrition (Jaarsveld et al., 2006)
Benjamin (2007) reported
that OFSP contains carbohydrates, β-carotene, vitamins
C and E, dietary fibre, minerals (K, Ca and Fe), protein, fat and cholesterol,
large amounts of thiamin (B1), riboflavin (B2), niacin (B3),
panthothenic acid (B5), pyridoxine (B6) and folic acid (B9),
and can lessen the effects of vitamin A deficiency as a result of its high β-carotene content. The significant
carotenoid present in OFSP is the β-carotene. Khoo et al. (2011) and Burri (2011) reported that β-carotene has the
highest vitamin A activity amid the several types of carotenoids. The amount of β-carotene contained in OFSP
depends on the variety of the sweet potato (Burri, 2011). Jaswir et al. (2011) reported that the total
β-carotene contained in OFSP is 20,000
μg.
OFSP is a beneficial root
crop, though it is underutilised in many parts of the world due to the
restricted information on the preparation of OFSP into other consumable
products (Assefa et al., 2007; Bezabih and Mengistu, 2011). Inadequate storage facilities and technology account for great loss of
OFSP after harvest (Ray and Ravi, 2005).
The liver is mainly responsible for drug metabolism, elimination
and detoxification of toxic substances (Mohammed et al., 2011).
Liver
damage-related diseases have become the ninth major cause of death worldwide
(Mohamed Saleem et al., 2010). Some
of the commonly suffered liver damage-related diseases include liver cirrhosis,
hepatitis and liver carcinoma. The major causes of some of these liver damage
related diseases are called hepatotoxins (chronic usage of some antibiotics,
paracetamol, some chemotherapeutic agents, thioacetamide and carbon tetrachloride)
which accounts for 50% of chronic liver diseases worldwide (Mcnally et al., 2006). Because of these commonly
used drugs which induce liver damage, stem cell derived hepatocytes should be
used to detect toxicity during drug development (Greenhough et al., 2012).
Various
hepatoprotective and antihepatotoxic agents have been effectively studied using
some of these hepatoxins; but over the years thioacetamide-induced liver damage
has been a more simple and effective model to study potential hepatoprotective
medicinal plants (Mohammed et al.,
2011). Thioacetamide causes liver damage through its metabolite
Thioacetamide-S-Oxide (TAASO) which increases the concentration of the
intracellular Ca+ thereby changing the cell permeability and causing
inhibition of the mitochondrial activity (Tasleem and Nadeem, 2013).
Hyperlipidaemia
is mainly characterized by elevation of serum concentration of several
lipoproteins (TC, TAG, LDL, and VLDL) and lipids. This is a major risk factor
for CVDs (cardiovascular diseases) and atherosclerosis which predispose to ischemic
heart disease (Hossain et al, 2011).
The pathology of atherosclerosis and other CVDs begins when the LDL becomes
oxidized in the vascular wall; the oxidation of LDL is as a result of the
production of the Reactive oxygen species (ROS) and Nitrogen species (NOS) by
the endothelial cells (Karimi et al.,
2013).
Hyperlipidaemia can be classified into primary
and secondary hyperlipidaemia. The primary hyperlipidemia is caused by genetic
mutation in the receptor protein, and it can be treated using hypolipidaemic
drugs while the secondary hyperlipidaemia is caused by metabolic disorders such
as diabetes, hypothyroidism, nephrotic syndrome, alcohol consumption, and it
can be reduced by treating the disease condition (Asija et al, 2014). The main factors which are responsible for hyperlipidaemia/dyslipidaemia
include genetic disorders, lifestyle and eating foods rich in cholesterol. Over
the years, a large number of synthetic drugs have been used for the treatment
of hyperlipidaemia but these drugs are linked with a lot of adverse effects.
Medicinal plants have been used to treat a lot of ailments due to the various
phytoconstituents contained in these plants, and they are also safer and more economical
than the synthetic drugs.
1.2 STATEMENT OF THE PROBLEM
Over
the years, there has been a consistent increase in the epidemiology of liver
damage related diseases despite the recent development of liver protective drugs;
some of these drugs are characterized with numerous side effects. Most of these
liver damage related diseases arises from toxicity of xenobiotic which
primarily affects the liver as a chief organ that carries out metabolism. The most
prevalent liver damage related diseases are liver cirrhosis, hepatitis and jaundice
which are responsible for a lot of death (Palanivel et al, 2008).
Despite
the availability of hyperlipidaemic drugs, the prevalence of hyperlipidemia (a
risk factor for coronary heart diseases) is still the leading cause of death in
most developing countries. Antihyperlipidaemic drugs accounts for numerous
adverse effects; severe muscle damage, liver damage, renal failure (Guyton and
Bays, 2007, Kobayashi et al, 2008,
Kiskac et al, 2013). Several
components of plants have been used to treat a lot of ailments around the world
with little or no adverse effects and some of these plant materials which
contain a lot of beneficial phytoconstituents are under-utilized and may
possess numerous pharmacological activities.
1.3 AIM
The
study is aimed at evaluating the hepatoprotective and antihyperlipidaemic
effect of orange flesh sweet potato (ipomea
batatas) juice in Sprague Dawley rat.
1.4 OBJECTIVES
The
objectives of the study were to:
1) Determine the phytochemical components present in orange
flesh potato juice.
2) Evaluate the chemical composition of OFSP using GC-MS
(Gas Chromatography and Mass Spectrometry).
3) Determine the effects of OFSP juice on biochemical (AST,
ALT, ALP, DB, TB) and haematological parameters (Hb, PCV, Platelets, RBC, TLC)
in the serum of thioacetamide-induced rats.
4) Evaluate the effect OFSP juice on the histopathology of the liver.
5) Assess the effect of OFSP juice on high fat
diet-induced hyperlipidaemia in rats using the lipid profile (CHOL, TAG ,HDL, LDL,
VLDL) and body weight parameters.
1.5 JUSTIFICATION OF STUDY
OFSP
has been reported by a lot of researchers to contain carbohydrates, β-carotene,
minerals, dietary fiber, vitamins, thiamin, riboflavin, niacin, pyridoxine and
folic acid. Some of these components and nutrients contained in OFSP can help
in the prevention and treatment of various diseases. Other varieties of sweet
potato have been reported to possess a lot of pharmacological activities; anti-ulcer,
anti-cancer, anti-inflammatory, immunomodulatory, hepatoprotective, anti-microbial,
anti-proliferative, anti-oxidant and anti-diabetic effects, but there are no
reported pharmacological potentials of OFSP. The present study is designed to investigate
some of the pharmacological potentials of OFSP so as to recommend its continued
use as foods, as well as processing it into more consumable products.
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