TABLE OF CONTENTS
TITLE PAGE – – – – – – – – I
APPROVAL PAGE – – – – – – – II
DEDICATION – – – – – – – – III
ACKNOWLEDGMENT – – – – – – IV
TABLE OF CONTENTS – – – – – – V
LIST OF TABLE – – – – – – – – VI
LIST OF FIGURE – – – – – – – – VII
ABSTRACT – – – – – – – – VIII
CHAPTER ONE
1.0 INTRODUCTION – – – – – – 1
CHAPTER TWO
2.0 LITERATURE REVIEW- – – – – – 4
2.1 TAXONOMY OF PLANT (DESMODIUM VELUTINUM) 4
2.2 LIVER- – – – – – – – – 10
2.3 WHAT THE LIVER DOES – – – – – 10
2.4 LIVER FUNCTION TEST/MARKER ENZYMES – 11
2.5 LIVER MARKERS ENZYMES (AST, ALT, ALP, GGT, LDH) 13
2.6 USES OF LIVER FUNCTION TEST – – – – 17
2.7 ATORVASTATIN – – – – – – 18
2.8 PHYTOCHEMICALS – – – – – – 20\
2.9 TYPES OF PHYTOCHEMICALS – – – – 21
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CHAPTER THREE
3.0 MATERIALS AND METHODS – – – – 24
3.1 IDENTIFICATION AND EXTRACTION OF DESMODIUM VELUTINUM PLANTS – – – – – – 26
3.2 PHYTOCHEMICAL ANALYSIS – – – – 27
3.3 EXPERIMENTAL ANIMAL MODEL – – – 31
3.4 COLLECTION OF BLOOD SAMPLES – – – 32
3.5 PROCEDURES FOR CARRYING OUT LIVER FUNCTION
TESTS – – – – – — – 33
CHAPTER FOUR
4.0 QUANTITATIVE ANALYSIS (PHYTOCHEMICAL COMPOSITION OF SAMPLES) – – – – 37
4.1 QUALITATIVE ANALYSIS (QUALITATIVE PHYTOCHEMISTRY) – – – – — – 37
4.2 LIVER FUNCTION TEST OF RATS FEED WITH VARIOUS SAMPLES – – – – – – – – 38
CHAPTER FIVE
5.0 DISCUSSION – – — – – – 39
5.2 CONCLUSION – – – – – – – 44
REFERENCES – – – – – – 46
LIST OF TABLES
TABLE I QUANTITATIVE ANALYSIS OF ETHANOL EXTRACT OF DESMODIUM VELUTINUM STEM
TABLE II QUALITATIVE ANALYSIS OF ETHANOL EXTRACT DESMODIUM
VELUTINUM STEM
TABLE III LIVER FUNCTION TESTS OF ALBINO WISTAR RATS FED WITH VARIOUS SAMPLES
LIST OF FIGURE
FIGURE I DESMODIUM VELUTINUM PLANT
ABSTRACT
Present investigations were carried out on the effect of ethanol extract of Desmodium velutinum stem on some liver enzymes of albino wistar rats fed with high fat from cow‟s brain. Twelve healthy albino wistar rats were divided into four groups. Group I rats were fed with balanced diet (normal feed, that is grower‟s mash) and tap water. Group II rats were fed with fat diet (cow‟s brain) (3.6g/day) alone. Group III rats received combined mixture of 70% fat diet (cow‟s brain) with 30% ethanol extract of Desmoduim elutinum stem (2.7g/day), Group IV rats were fed with fat diet (cow‟s brain) and were administered with a drug atorvastatin (5mg/rat/day). The rats were sacrificed at the end of the experimental (two weeks) period. The plant extract was screened for its phytochemical constituents, using standard procedures. Phytochemical screening revealed the presence of tannins, flavonoids, saponins and alkaloids. The liver function tests was assessed by measuring the total bilirubin, total serum protein, direct and indirect bilirubin and liver enzymes such as Alanine amino transferase (ALT), Aspartate amino transferase (AST), Alkaline phosphate (ALP), Gamma-glutamyl transferase (GGT) on the liver of the albino wistar rat. The serum, ALT, ALP, AST and bilirubin concentration of the albino wistar rat increased significantly (P>0.05) compared to their total protein concentration. The result obtained indicate that the ethanol extract of Desmodium Velutinum stem possess hepatoprotective activity on the fatty liver of albino wistar rat fed with fat from cow‟s brain. The studied plant showed variable amount of phytochemicals including flavonoid, alkaloid, saponin, tannins, terpenoid which were determined quantitatively using literature methods. The significant effect of the Desmodium velutinum stem ethanol extract was compared with standard drug, atorvastatin. The result in the present study suggest that the ethanol extract of Desmodium velutinum stem can be used in treating liver diseases.
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CHAPTER ONE
INTRODUCTION
The liver is the largest and most complex internal organ in the body. It plays an important role in the maintenance of internal environment through its multiple and diverse functions. Liver is involved in several vital functions such as metabolism, secretion and storage. Hepatitis or inflammatory disorder involves inflammation and change to the hepatocytes. Hepatitis is one of the most prevalent diseases in the world. Every year 18,000 people had been reported to die due to liver cirrhosis caused by viral hepatitis (Wang et al., 2008). Ectopic fat storage occurs in obesity, particularly in the liver leading to a condition termed non alcoholic fatty liver disease (NAFLD) characterized by varying degree of liver injury that progresses from steatosis to tratohepatitis, fibrosis and necrosis. Due to its prominent association with insulin resistance/obesity, NAFLD is regarded as the hepatic manifestation of metabolic syndrome. Liver has great capacity to detoxicate toxic substance and synthesizes useful principles. Therefore damage to liver inflicted by hepatotoxic agents is of grave consequences. Experimental studies have reported that animal fed a high fat diet (HFD) for more than two months develop weight, hyperlipidemic, hyperglycemia, oxidative stress and insulin resistance (IR). Besides, consumption of a calorie-rich diet results in lipid accumulation, excess production of inflammatory cytokines and macrophage infiltration that favours the
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progression of liver disease. Many medicinal plant/indigenous plant have been mentioned and well established as hepatoprotective agents.
Desmodium velutinum is a medicinal plant around the world. The extract of Desmodium velutinum showed significant anti-pyretic activity on experimental rats used. Desmodium velutinum is very rich in alkaloids and related amino compounds. It is a source of flavonoid, saponins and pharmacological active agent useful in the treatment of aches and pains. It also showed that one of the uses is for erectile enhancing property. Desmodium velutinum being one of the important medicinal plant in the world may have effect on the liver. However limited studies are available on the effect of Desmodium velutinum treatment on hepatoprotective effect of its extract on high fat diet fed animal. In this regard, the present study was aimed to investigate the effect of ethanol extract of Desmodium velutinum stem on liver function tests on albino wistar rats fed with high fat from cow‟s brain related to different hepatoprotective effect by comparing it to a known drug, atorvastatin treatment which is used to lower the level of cholesterol in blood. Since changes in the concentration and composition of plasma lipid and lipoproteins occur frequently in liver disease, these findings may be useful in following the clinical course of patient with liver diseases of various causes.
Liver disease is associated with characteristic changes in the lipid composition of the surface coat of plasma lipoprotein particles. Cholesterol accumulate as hepatic
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secretion of lecithin cholesterol acyltransferase decreases, such abnormal circulating lipoproteins tend to induce corresponding changes in cell membrane lipid composition. Studies in both human and experimental liver disease confirm that this does occur and that it is wide-spread. Low density lipoproteins, which are rich in cholesterol can also be taken up by the liver or extrahepatic tissues by a receptor mediated endocytosis that specially recognizes apolipoproteins. It can be predicted that extra hepatic membrane dysfunction might be a general feature of severe liver disease. It is proposed that many of the cellular disturbances and metabolic abnormalities accompanying hepatic disease result from, or are exacerbated by lipoprotein induced changes in membrane lipid composition and function. Hence Desmodium velutinum may be a source of a pharmacological active agent useful in the treatment of high level cholesterol in the blood which has got both liver protecting and nutritional value.
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