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Phd Thesis- Phyllis E. A. Dadzeasah.Pdf SAFETY EVALUATION AND HEPATOPROTECTIVE ACTIVITY OF THE AQUEOUS STEM BARK EXTRACT OF Spathodea campanulata A THESIS SUBMITTED IN FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY In the Department of Pharmacology, Faculty of Pharmacy and Pharmaceutical Sciences by PHYLLIS ELSIE ABAIDUWA DADZEASAH KWAME NKRUMAH UNIVERSITY OF SCIENCE AND TECHNOLOGY, KUMASI JUNE, 2012 DECLARATION The experimental work described in this thesis was carried out at the Department of Pharmacology, KNUST. This work has not been submitted for any other degree. Phyllis Elsie Abaiduwa Dadzeasah ………………… ……………….. ID: 20066532 (Signature) (Date) Index No: PG2743908 Rev. Prof. Charles Ansah ………………… ……………….. (Supervisor) (Signature) (Date) …………………………. ………………….. ........................ (Head of Department) (Signature) (Date) ii ABSTRACT Spathodea campanulata, a widely used traditional African medicinal plant for skin diseases and stomach aches has not been scientifically assessed for toxicity and hepatoprotection. The aqueous extract of the stem bark of the plant Spathodea campanulata (SCE) was therefore evaluated for safety and potential hepatoprotective activity in rodents. The extract (1250, 2500, 5000 mg/kg, p.o) administered to rats and mice over a 24-hour period did not show any signs of toxicity or mortality, suggesting that the oral LD50 of the aqueous extract (in rats and mice) was beyond 5000 mg/kg. Daily administration (1250, 2500, 5000 mg/kg; p.o) of extract for 14 days did not cause any changes in behavior or alterations in haematological parameters of the animals. Serum biochemical analysis in rats showed a rise in alkaline phosphatase (ALP) (339.50±90.50 in the control to 582.5±28.50 U/L in the highest dose of 5000 mg/kg) and a decrease in Na+ concentration (147.3±1.856 mmol/L in the control to 135.0±1.00 in the 5000mg/kg dose), but no change in other parameters. These observations did not reflect in the histopathology of the liver and kidneys of treated animals. However, the diuretic test on the extract was positive. Urine output, Na+ and K+ concentration in urine were increased significantly by the administration of extract for seven days. In hepatoprotective studies, rats were pre-treated with 625 mg/kg , 1250 mg/kg, 2500 mg/kg p.o for 4 days before intoxication with carbon tetrachloride (CCl4) (1 ml/kg, 20 % in liquid paraffin, p.o) or mice and rats with 100 mg/kg, 300 mg/kg, 625 mg/kg for 4 days before intoxication with paracetamol or aflatoxin B1 (AFB1) respectively. Generally, the administration of these hepatotoxic agents caused an increase in liver weights which were prevented or restored by pre-treatment with extract in both rats and mice. The aspartate transaminase (AST), alanine transaminase (ALT), gamma glutamyl transpeptidase (GGT), bilirubin-induced by these hepatotoxic agents (CCl4, parcetamol and AFB1) were significantly reduced with SCE treatment. Additionally, administration of CCl4 daily for 5 days, followed by treatment with 100, 300, and 625 mg/kg p.o daily for 3days (curative) decreased profoundly the AST, ALT and GGT levels induced by CCl4. These results iii correlated well with the histopathological studies observed in the photomicrographs of liver for treated and control groups. Treatment with the extract decreased the extent of fatty liver and necrosis caused by the hepatotoxic agents. Lipid peroxidation measured as TBARS was elevated by CCl4 but this was reversed by treatment with the extract at all dose levels. Again, pretreatment with SCE (625 mg/kg) before hepatotoxicity by CCl4 or paracetamol restored the decrease in super oxide dismutase (SOD) and glutathione peroxidase (GPx) activity caused by the hepatotoxin. This decrease was more profound after paracetamol intoxication than it was after CCl4 intoxication. The extract (625 mg/kg) also showed strong enzyme inhibition when it reduced the total cytochrome P450 in both mice and rats when given for seven consecutive days. This decrease was also reflected in the presence of CCl4 and paracetamol. The preliminary phytochemistry which showed the presence of tannins, sterols and reducing sugars as well as the in vitro testing which gave positive results for reducing power and total phenolic content also support the activity of the plant extract in interference with hepatotoxicity. Collectively, the results indicate that the aqueous extract of Spathodea campanulata is fairly nontoxic and may exhibit hepatoprotective activity at lower doses by enhancing antioxidant protection in the cell and inhibiting total cytochrome P450 hence interfering with bioactivation of hepatotoxic agents. iv ACKNOWLEDGEMENT I am grateful to the almighty God for seeing me through the programme. Surely, it has been His grace. I also wish to express my profound gratitude to my supervisor, Rev. Prof. Charles Ansah of the Department of Pharmacology, KNUST, for supervision, particularly his suggestions, criticisms, support and mentorship which saw to the realization of this project. I am also grateful to Mr. Thomas Ansah and indeed the entire technical staff of the Department of Pharmacology, KNUST and Mr. Emmanuel Assiamah of the Department of Histology, Komfo Anokye Teaching Hospital, for their technical support and the whole staff and colleague students in the department of Pharmacology for their support. Finally, my heartfelt gratitude goes to my family and friends for their sacrifice and support. Mum and Dad, this was much your dream as it was mine; Philip, your encouragement and emotional support can never be overlooked. v TABLE OF CONTENTS DECLARATION .......................................................................................................................... II ABSTRACT ................................................................................................................................ III ACKNOWLEDGEMENT .......................................................................................................... V TABLE OF CONTENTS .......................................................................................................... VI LIST OF TABLES ................................................................................................................. XVII LIST OF FIGURES ............................................................................................................. XVIII ABBREVIATIONS .............................................................................................................. XXIII CHAPTER 1 .................................................................................................................................. 1 GENERAL INTRODUCTION .................................................................................................. 1 1.1 OVERVIEW ....................................................................................................................... 2 1.2 GLOBAL USE OF HERBAL REMEDIES ................................................................................... 2 1.3 THE PLANT SPATHODEA CAMPANULATA ............................................................. 4 1.3.1 Medicinal Uses ........................................................................................................... 6 Traditional uses ................................................................................................................... 6 1.3.2 Biological Activities of Spathodea campanulata ....................................................... 7 Molluscicidal Activity ........................................................................................................ 7 vi Hypoglycaemic Activity ..................................................................................................... 7 Antimalarial Activity .......................................................................................................... 7 Anti-inflammatory and antioxidant properties................................................................... 8 Antibacterial Activity ......................................................................................................... 8 Wound healing Activity...................................................................................................... 8 1.3.3 Some Identified Chemical Constituents of Spathodea campanulata ........................ 8 1.3.4 Non-medicinal uses .................................................................................................. 12 1.4 SAFETY EVALUATION OF MEDICINAL PLANTS .................................................. 13 1.4.1 Factors influencing Toxicity .................................................................................... 14 1.4.2 Assessment of Toxicity .............................................................................................. 15 Acute Toxicity .................................................................................................................. 15 Chronic Toxicity ............................................................................................................... 16 1.4.3 Target organ Toxicity ............................................................................................... 17 Blood Toxicity .................................................................................................................. 17 Kidney Toxicity ................................................................................................................ 18 Liver Toxicity ..................................................................................................................
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