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ANALGESIC and ANTI-INFLAMMATORY ACTIVITIES of METHANOL EXTRACTS of Pistacia Aethiopica (Kokwaro) and Warbugia Ugandensis (Sprague) in MICE MODELS

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ANALGESIC and ANTI-INFLAMMATORY ACTIVITIES of METHANOL EXTRACTS of Pistacia Aethiopica (Kokwaro) and Warbugia Ugandensis (Sprague) in MICE MODELS ANALGESIC AND ANTI-INFLAMMATORY ACTIVITIES OF METHANOL EXTRACTS OF Pistacia aethiopica (Kokwaro) and Warbugia ugandensis (Sprague) IN MICE MODELS IRERI MOSES MUNENE (BSc.) I56/CE/26169/2014 A THESIS SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE AWARD OF THE DEGREE OF MASTER OF SCIENCE IN BIOTECHNOLOGY IN THE SCHOOL OF PURE AND APPLIED SCIENCES OF KENYATTA UNIVERSITY SEPTEMBER, 2019 ii DECLARATION I, IRERI MOSES MUNENE, duly declare that this thesis is my original work and has not been presented for a degree in any other university or for any other award. IRERI MOSES MUNENE (BSc.) I56/CE/26169/2014 Signature ___________________________Date ___________________________ Supervisors We confirm that the work reported in this thesis was carried out by the candidate under our supervision. Signature ___________________________Date ___________________________ Prof Eliud NM Njagi Department of Biochemistry, Microbiology and Biotechnology Kenyatta University P.O. Box 43844-00100 Nairobi, Kenya Signature ___________________________Date ___________________________. Dr Mathew Piero Ngugi Department of Biochemistry, Microbiology and Biotechnology Kenyatta University P.O. Box 43844-00100 Nairobi, Kenya iii DEDICATION This thesis is dedicated to my mum, Rose Mary Njeru, for her love, relentless prayers and immense support towards my education. iv ACKNOWLEDGEMENTS I am greatly indebted to Kenyatta University for giving me an opportunity to further my education and get to be a member of the alumni. I owe sincere gratitude to my supervisors Prof Eliud NM Njagi and Dr Mathew Piero Ngugi for tremendous guidance, inspiration and support that greatly facilitated completion of this research study and left me with a great sense of fulfillment and achievement which will continue to influence my work. Your valuable mentorship is greatly cherished. I also owe gratitude to the following people for their enormous support. Thanks to the entire staff of Biochemistry and Biotechnology department for all the assistance. The following people deserve special mention, Daniel Gitonga and James Ngunjiri for technical assistance. To John K. Mwonjoria, James Kimani, Samson Koech, Veronica Sindani, Peter Nthiga, Lawrence Alaro and Berrick Moturi for your encouragement and assistance which greatly made my work a success. To my mum, Rosemary Njeru, your financial support and prayers made completion of this work possible. Thank you for believing in me. To Justin Fundi and Caroline Wanja your encouragement is greatly appreciated. Above all, I owe great reverence and gratitude to the Almighty God, for granting me strength, robust health and sound mind to accomplish this project. From Him is all knowledge, understanding, power and wisdom, glory be to His holy name. Lastly, to all who contributed to the success of my work, may the almighty God bless them abundantly. v TABLE OF CONTENTS DECLARATION ....................................................................................................... ii DEDICATION .......................................................................................................... iii ACKNOWLEDGEMENTS ..................................................................................... iv TABLE OF CONTENTS .......................................................................................... v LIST OF TABLES .................................................................................................. viii LIST OF FIGURES .................................................................................................. ix LIST OF APPENDICES ........................................................................................... x ABBREVIATIONS AND ACRONYMS ................................................................ xi ABSTRACT ............................................................................................................. xii CHAPTER ONE ........................................................................................................ 1 INTRODUCTION ..................................................................................................... 1 1.1 Background information ........................................................................................ 1 1.2 Problem statement and justification ...................................................................... 4 1.3 Null Hypotheses .................................................................................................... 6 1.4 Objectives .............................................................................................................. 6 1.4.1 General objective ............................................................................................ 6 1.4.2 Specific objectives .......................................................................................... 6 CHAPTER TWO ....................................................................................................... 7 LITERATURE REVIEW ......................................................................................... 7 2.1 Physiological and biochemical basis of pain and inflammation ........................... 7 2.1.1 Pain ................................................................................................................. 7 2.1.2 Inflammation ................................................................................................ 11 2.2 Conventional management of pain and inflammation......................................... 17 2.3 Herbal management of pain and inflammation ................................................... 19 vi 2.4 Models used in pain and inflammation studies ................................................... 21 2.4.1 Models used in pain studies .......................................................................... 21 2.4.1.1 Models using chemical stimulus ........................................................... 21 2.4.1.2 Models using thermal stimuli ................................................................ 23 2.4.2 Models used in inflammation studies ........................................................... 24 2.4.2.1 Carrageenan induced paw edema .......................................................... 24 2.4.2.1 Egg albumin induced edema ................................................................. 25 2.5 Plants used in this study ...................................................................................... 26 2.5.1 Warbugia ugandensis Sprague ..................................................................... 26 2.5.1.1 Classification and distribution ............................................................... 26 2.5.1.2 Plant description .................................................................................... 26 2.5.1.3 Medicinal use ......................................................................................... 27 2.5.2 Pistacia aethiopica Kokwaro ....................................................................... 28 2.5.2.1 Classification and distribution ............................................................... 28 2.5.2.2 Plant description .................................................................................... 28 2.5.2.3 Medicinal use ......................................................................................... 29 CHAPTER THREE ................................................................................................ 30 MATERIALS AND METHODS ............................................................................ 30 3.1 Collection and preparation of plant materials ..................................................... 30 3.2 Extraction ............................................................................................................ 30 3.3 Experimental design ............................................................................................ 31 3.3.1 Laboratory animals ....................................................................................... 31 3.4 Bioscreening ........................................................................................................ 31 3.4.1 Determination of analgesic activities ........................................................... 31 3.4.2 Determination of anti-inflammatory activities ............................................. 33 vii 3.5 Qualitative Phytochemical Screening ................................................................. 36 3.5.1 Test for saponins ........................................................................................... 36 3.5.2 Test for alkaloids .......................................................................................... 36 3.5.3 Test for terpenoids ........................................................................................ 36 3.5.4 Test for flavonoids ........................................................................................ 37 3.5.5 Test for cardiac glycosides ........................................................................... 37 3.5.6 Test for steroids ............................................................................................ 37 3.5.7 Test for phenols ............................................................................................ 37 3.5.8 Test for tannins ............................................................................................. 38 3.6 Data Management and Statistical Analysis ......................................................... 38 CHAPTER FOUR ..................................................................................................
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