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EFFECT OF AQUEOUS EXTRACT OF Lawsonia inermis LEAVES ON THE LIVER FOLLOWING ACUTE ETHANOL-INDUCED HEPATIC DAMAGE IN WISTAR RATS

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

INTRODUCTION

Background of the Study

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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