University of Macau / Institute of Chinese Medical Sciences

University of Macau / Institute of Chinese Medical Sciences

University of Macau / Institute of Chinese Medical Sciences

University of Macau / Institute of Chinese Medical Sciences

Lam, Ming Yiu. / "September 2011." / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 149-169). / Abstracts in English and Chinese. / Thesis / Assessment Committee --- p.ii / Abstract (English) --- p.iii / Abstract (Chinese) --- p.vi / Acknowledgements --- p.viii / Table of contents --- p.x / List of figures --- p.xvi / List of tables --- p.xix / Abbreviations --- p.xx / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Cerebral stroke --- p.1 / Chapter 1.2 --- Epidemiology --- p.2 / Chapter 1.3 --- Risk factors and symptoms --- p.5 / Chapter 1.3.1 --- Non-modifiable risks --- p.5 / Chapter 1.3.2 --- Modifiable risks --- p.6 / Chapter 1.3.3 --- Symptoms --- p.9 / Chapter 1.4 --- Mechanisms of cell injury --- p.10 / Chapter 1.4.1 --- Energy failure and loss of ionic homeostasis --- p.10 / Chapter 1.4.2 --- Excitotoxicity and calcium-modulated cell damage --- p.11 / Chapter 1.4.3 --- Oxidative stress --- p.13 / Chapter 1.4.4 --- Inflammation --- p.16 / Chapter 1.4.5 --- Apoptosis --- p.18 / Chapter 1.5 --- Current treatment of ischemia --- p.19 / Chapter 1.6 --- Chinese herbal medicine --- p.21 / Chapter 1.6.1 --- Traditional Chinese medicine theory on stroke --- p.21 / Chapter 1.6.2 --- Danshen --- p.22 / Chapter 1.6.3 --- Gegen --- p.25 / Chapter 1.6.4 --- Danshen-Gegen formula --- p.28 / Chapter 1.7 --- Aim of study --- p.30 / Chapter Chapter 2 --- General methodology --- p.31 / Chapter 2.1 --- Induction of transient focal cerebral ischemia by middle cerebral artery occlusion (MCAO) --- p.31 / Chapter 2.1.1 --- Intraluminal filament production --- p.32 / Chapter 2.1.2 --- Cerebral blood flow measurement by laser Doppler flowmetry --- p.33 / Chapter 2.1.3 --- Middle cerebral artery occlusion --- p.35 / Chapter 2.2 --- Neurological scoring --- p.38 / Chapter 2.3 --- Brain infarction measurement by triphenyltetrazolium chloride (TTC) staining --- p.40 / Chapter 2.4 --- Statistical analyses --- p.42 / Chapter Chapter 3 --- Preparation of herbal medicine --- p.43 / Chapter 3.1 --- Authentication of Chinese herbs --- p.43 / Chapter 3.1.1 --- Morphological authentication --- p.43 / Chapter 3.1.2 --- Chemical authentication using thin layer chromatography --- p.44 / Chapter 3.1.2.1 --- Danshen --- p.44 / Chapter 3.1.2.2 --- Gegen --- p.48 / Chapter 3.2 --- Danshen-Gegen (DG) extract preparation --- p.50 / Chapter 3.3 --- Chemical analysis of DG extract --- p.51 / Chapter 3.3.1 --- TLC --- p.51 / Chapter 3.3.2 --- HPLC --- p.54 / Chapter 3.4 --- Conclusion --- p.57 / Chapter Chapter 4 --- Protective effect of DG extract on cerebral ischemia --- p.58 / Chapter 4.1 --- Introduction --- p.58 / Chapter 4.1.1 --- Different models of ischemia --- p.58 / Chapter 4.1.2 --- Anti-oxidative enzymes in cerebral ischemia --- p.61 / Chapter 4.1.2.1 --- Superoxide dismutase (SOD) --- p.61 / Chapter 4.1.2.2 --- Catalase --- p.62 / Chapter 4.1.2.3 --- Glutathione peroxidase (GPX) --- p.62 / Chapter 4.2 --- Materials and methods --- p.64 / Chapter 4.2.1 --- "DG extract treatment, neurological deficit and brain infarction" --- p.64 / Chapter 4.2.2 --- Anti-oxidative enzymes activity determination --- p.65 / Chapter 4.2.2.1 --- Treatment with DG extract and induction of cerebral ischemia --- p.65 / Chapter 4.2.2.2 --- Extraction of enzymes from the brain --- p.66 / Chapter 4.2.2.3 --- Determination of protein concentration --- p.66 / Chapter 4.2.2.4 --- Assay kits --- p.67 / Chapter 4.3 --- Results --- p.70 / Chapter 4.4 --- Discussion --- p.80 / Chapter 4.4.1 --- Neurological score and percentage brain infarction --- p.80 / Chapter 4.4.2 --- Anti-oxidative enzyme induction --- p.82 / Chapter Chapter 5 --- Behavioral assessment using the shuttle box avoidance test on rats suffering from cerebral ischemia: effect of DG extract treatment --- p.86 / Chapter 5.1 --- Introduction --- p.86 / Chapter 5.1.1 --- Behavioral tests --- p.86 / Chapter 5.1.2 --- Theory of the test --- p.90 / Chapter 5.2 --- Materials and methods --- p.91 / Chapter 5.2.1 --- DG extract treatment --- p.91 / Chapter 5.2.2 --- Shuttle box training and MCAO --- p.92 / Chapter 5.2.3 --- Shuttle box testing --- p.96 / Chapter 5.2.4 --- Neurological score and brain infarction --- p.96 / Chapter 5.3 --- Results --- p.97 / Chapter 5.3.1 --- Shuttle box performance --- p.97 / Chapter 5.3.2 --- Neurological score --- p.105 / Chapter 5.3.3 --- Brain infarction --- p.109 / Chapter 5.4 --- Discussion --- p.112 / Chapter 5.4.1 --- The shuttle box protocol --- p.112 / Chapter 5.4.2 --- Shuttle box performance --- p.114 / Chapter 5.4.2.1 --- Pretreatment groups --- p.114 / Chapter 5.4.2.2 --- Pre + post treatment groups --- p.115 / Chapter 5.4.2.3 --- Comparison of pretreatment and pre + post treatment groups --- p.116 / Chapter 5.4.3 --- Neurological score --- p.117 / Chapter 5.4.4 --- Brain infarction --- p.118 / Chapter 5.4.5 --- Conclusion --- p.119 / Chapter Chapter 6 --- Anti-blood coagulation effect of DG extract --- p.121 / Chapter 6.1 --- Introduction --- p.121 / Chapter 6.2 --- Materials and methods --- p.125 / Chapter 6.2.1 --- Treatment with DG extract and warfarin --- p.125 / Chapter 6.2.2 --- Tail bleeding time and volume --- p.126 / Chapter 6.2.3 --- Prothrombin time --- p.127 / Chapter 6.2.4 --- Platelet aggregation --- p.127 / Chapter 6.3 --- Results --- p.128 / Chapter 6.4 --- Discussion --- p.138 / Chapter Chapter 7 --- General discussion --- p.141 / Chapter 7.1 --- General discussion and conclusion --- p.141 / Chapter 7.2 --- Clinical significance of the study --- p.145 / Chapter 7.3 --- Limitations of the study --- p.146 / Chapter 7.4 --- Future work --- p.147 / References --- p.149 / Publications --- p.170

University of Macau / Institute of Chinese Medical Sciences

University of Macau / Institute of Chinese Medical Sciences

Ng, Chun Fai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 134-150). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.vii / Table of Contents --- p.ix / Abbreviations --- p.xii / List of Figures --- p.xv / List of Tables --- p.xviii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- "Introduction to Cardiovascular Disease, Hypertension and Atherosclerosis" --- p.1 / Chapter 1.1.1 --- Cardiovascular Disease --- p.1 / Chapter 1.1.2 --- Hypertension --- p.2 / Chapter 1.1.2.1 --- Background --- p.2 / Chapter 1.1.2.2 --- Causes of Hypertension --- p.3 / Chapter 1.1.2.3 --- Current Western Management and Medication --- p.6 / Chapter 1.1.3 --- Atherosclerosis --- p.9 / Chapter 1.1.3.1 --- Background --- p.9 / Chapter 1.1.3.2 --- Pathogenesis of Atherosclerosis --- p.10 / Chapter 1.1.3.3 --- Current Western Treatment and Medication --- p.12 / Chapter 1.2 --- Selection and Introduction of Current Chinese Medicine Formula --- p.16 / Chapter 1.2.1 --- Cardiac Syndrome in Traditional Chinese Medicine --- p.16 / Chapter 1.2.2 --- Traditional Chinese Medicine as an Complementary or Alternative Medicine --- p.17 / Chapter 1.2.3 --- Selection of TCM Formula from Pharmacopoeia --- p.18 / Chapter 1.2.3.1 --- Compound Formula --- p.18 / Chapter 1.2.4 --- Introduction to Constitutional Herbal Medicine --- p.19 / Chapter 1.2.4.1 --- Danshen (Radix Salviae miltiorrhizae) --- p.19 / Chapter 1.2.4.2 --- Gegen (Radix Puerariae lobatae) --- p.20 / Chapter 1.2.4.3 --- Yanhusuo (Rhizoma Corydalis) --- p.21 / Chapter 1.2.4.4 --- Composition of the Final Formula Used in the Present Study --- p.21 / Chapter 1.2.5 --- Previous work on Danshen-Gegen Formula and its limitations --- p.22 / Chapter 1.3 --- Objectives of the Present Study --- p.25 / Chapter 1.3.1 --- Research Plan --- p.26 / Chapter Chapter 2 --- Experimental Design and General Methodology --- p.27 / Chapter 2.1 --- Source and Authentication of Raw Herbs --- p.27 / Chapter 2.2 --- Materials --- p.29 / Chapter 2.3 --- Ethical Approval --- p.31 / Chapter 2.4. --- General Methods --- p.32 / Chapter 2.4.1 --- Blood Pressure Measurement --- p.32 / Chapter 2.4.2 --- Blood Profile Measurement --- p.33 / Chapter 2.4.3 --- Vascular Reactivity Studies --- p.36 / Chapter 2.5 --- Statistical Analysis --- p.38 / Chapter Chapter 3 --- Anti-hypertensive Studies of Danshen-Gegen Formula on Rat --- p.39 / Chapter 3.1 --- Introduction --- p.39 / Chapter 3.1.1 --- In vivo Anti-Hypertensive Studies --- p.39 / Chapter 3.1.1.1 --- Spontaneously Hypertensive Rat (SHR) --- p.40 / Chapter 3.1.1.2 --- Tail-cuff Blood Pressure Measurement --- p.41 / Chapter 3.1.2 --- Detailed Underlying Mechanistic Studies --- p.42 / Chapter 3.1.2.1 --- Nitric Oxide-mediated Vasodilation --- p.42 / Chapter 3.1.2.2 --- Prostacyclin-mediated Vasodilation --- p.43 / Chapter 3.1.2.3 --- Hyperpolarization-mediated Vasodilation --- p.43 / Chapter 3.1.2.4 --- Endothelium-dependent/-independent Vasodilation --- p.46 / Chapter 3.1.3 --- Long Term Underlying Mechanistic Studies --- p.48 / Chapter 3.2 --- Methods --- p.49 / Chapter 3.2.1 --- In vivo Anti-Hypertensive Studies --- p.49 / Chapter 3.2.2 --- Detailed Underlying Mechanistic Studies --- p.51 / Chapter 3.2.3 --- Long Term Underlying Mechanistic Studies --- p.53 / Chapter 3.2.4 --- Statistical analysis --- p.56 / Chapter 3.3 --- Results --- p.58 / Chapter 3.3.1 --- In vivo Anti-Hypertensive Studies --- p.58 / Chapter 3.3.1.1 --- Preventive Effect in Hypertension --- p.58 / Chapter 3.3.1.2 --- Therapeutic Effect in Hypertension --- p.62 / Chapter 3.3.2 --- Detailed Underlying Mechanistic Studies --- p.66 / Chapter 3.3.2.1 --- DG extract-induced Vasodilation --- p.66 / Chapter 3.3.2.2 --- Endothelium-independent Vasodilation --- p.67 / Chapter 3.3.2.3 --- Nitric Oxide-mediated Vasodilation --- p.68 / Chapter 3.3.2.4 --- Prostacyclin-mediated Vasodilation --- p.69 / Chapter 3.3.2.5 --- Hyperpolarization-mediated Vasodilation --- p.70 / Chapter 3.3.3 --- Long Term Underlying Mechanistic Studies --- p.74 / Chapter 3.4 --- Discussion --- p.79 / Chapter Chapter 4 --- Anti-atherosclerosis Studies of Danshen-Gegen Formula in Rabbits --- p.89 / Chapter 4.1 --- Introduction --- p.89 / Chapter 4.1.1 --- Intima-Media Thickening --- p.89 / Chapter 4.1.2 --- Effect of High Cholesterol Diet in Rabbit --- p.90 / Chapter 4.1.3 --- Thiobarbituric Acid Reactive Substances --- p.91 / Chapter 4.2 --- Methods --- p.93 / Chapter 4.2.1 --- Pilot Study for Establishment of Experimental Protocol --- p.93 / Chapter 4.2.2 --- Effect of DG extract on Intima-media Thickening --- p.97 / Chapter 4.2.3 --- Statistical analysis --- p.99 / Chapter 4.3 --- Result --- p.100 / Chapter 4.3.1 --- Study of the Anti-atherosclerosis Effect of DG extract - First Run --- p.100 / Chapter 4.3.2 --- Study of the Anti-atherosclerosis Effect of DG extract - Second Run --- p.108 / Chapter 4.4 --- Discussion --- p.117 / Chapter Chapter 5 --- General Discussion and Conclusion --- p.122 / Chapter 5.1 --- Significance of the Study --- p.122 / Chapter 5.2 --- Limitations and Future work --- p.127 / Chapter 5.3 --- Clinical Implication of the Use of the DG Preparations for Patients with CVD --- p.132 / References --- p.134

published_or_final_version / Biological Sciences / Doctoral / Doctor of Philosophy

published_or_final_version / Chemistry / Master / Master of Philosophy