Investigation of in vitro and in vivo effects of raloxifene on the pulmonary and systemic vascular circulations.

Chan Yau Chi.Thesis (M.Phil.)--Chinese University of Hong Kong, 2005.Includes bibliographical references (leaves 157-177).Abstracts in English and Chinese.ContentsDeclaration --- p.iAcknowledgement --- p.iiAbbreviations --- p.iii-ivAbstract in English --- p.v-viiiAbstract in Chinese --- p.ix-xiContents --- p.xii-xviChapter CHAPTER I - --- IntroductionChapter 1.1. --- Selective Estrogen Receptor Modulators (SERMs) --- p.1Chapter 1.1.1. --- Raloxifene --- p.6Chapter 1.2. --- Mechanisms of Action of SERMs in Vascular System --- p.7Chapter 1.2.1. --- Estrogen --- p.7Chapter 1.2.2. --- Estrogen Receptors (ERs) --- p.8Chapter 1.2.3. --- General Mechanisms of Action of SERMs --- p.13Chapter 1.2.4. --- Actions of Raloxifene --- p.14Chapter 1.3. --- Effects of SERMs in Cardiovascular System --- p.14Chapter 1.3.1. --- Effects of SERMs on Endothelial Function --- p.15Chapter 1.3.2. --- Effects of SERMs on Vascular Smooth Muscle --- p.17Chapter 1.4. --- Effects of Raloxifene on Vascular Circulations --- p.18Chapter 1.4.1. --- Effects of Raloxifene on Systemic Circulation --- p.18Chapter 1.4.1.1. --- Preclinical Data --- p.18Chapter 1.4.1.1.1. --- Effects on Serum Lipids --- p.18Chapter 1.4.1.1.2. --- Effects on Inflammation Markers and Blood Coagulation --- p.19Chapter 1.4.1.1.3. --- Antioxidative Effects --- p.19Chapter 1.4.1.1.4. --- Effects on Nitric Oxide and Endothelial Function --- p.19Chapter 1.4.1.1.5. --- Effects on Vascular Smooth Muscle --- p.20Chapter 1.4.1.1.6. --- "Vascular Injury, Atherosclerosis and Ischaemia-Reperfusion Injury" --- p.20Chapter 1.4.1.2. --- Clinical Studies - Effects in Post-Menopausal Women --- p.21Chapter 1.4.1.2.1. --- "Effects on Serum Lipids, Lipoproteins and Triglycerides" --- p.21Chapter 1.4.1.2.2. --- Effects on Inflammation Markers and Homocysteine --- p.22Chapter 1.4.1.2.3. --- Effects on Coagulation Markers --- p.23Chapter 1.4.1.2.4. --- Effects on Endothelial Function --- p.23Chapter 1.4.1.2.5. --- Cardiovascular Events --- p.23Chapter 1.5. --- Myogenic Response and Vascular System --- p.24Chapter 1.5.1. --- Initiation and Development of Myogenic Response --- p.25Chapter 1.5.2. --- Regulation of Myogenic Response --- p.26Chapter 1.5.2.1. --- 20-hydroxyeicosatetraenoic acid (20-HETE) --- p.26Chapter 1.5.2.2. --- "Protein Kinase C, Rho/Rho-Kinase, and Tyrosine Kinase" --- p.27Chapter 1.5.3. --- Myogenic Response and Endothelium --- p.31Chapter 1.5.4. --- Estrogen and Myogenic Tone --- p.31Chapter 1.6. --- Objectives of the Present Study --- p.32Chapter CHAPTER II - --- Methods and MaterialsChapter 2.1. --- Tissue and Cell Preparation --- p.34Chapter 2.1.1. --- Vessel Preparation --- p.34Chapter 2.1.2. --- Removal of a Functional Endothelium --- p.36Chapter 2.2. --- Myograph and Pressure Myograph Setups --- p.36Chapter 2.2.1. --- Myograph 一 Isometric Tension Measurement --- p.36Chapter 2.2.2. --- Pressure Myograph - Isobaric Diameter Measurement --- p.37Chapter 2.3. --- Intracellular [Ca2+] Measurement in Vascular Smooth Muscle --- p.42Chapter 2.4. --- Chronic Raloxifene Therapyin Spontaneously Hypertensive Rats (SHRs) and Wistar-Kyoto Rats (WKYs) --- p.42Chapter 2.4.1. --- Surgical Procedure - Raloxifene Tubing Insertion --- p.42Chapter 2.4.2. --- "Body Weight, Mean Arterial Blood Pressure and Uterine Weight" --- p.42Chapter 2.4.3. --- Measurement of Raloxifene Tubing Consumption --- p.43Chapter 2.4.4. --- Effect of Chronic Raloxifene Treatment on Artery Reactivity --- p.43Chapter 2.5. --- Ovariectomy and Chronic Raloxifene Therapyin Syrian Golden Hamsters --- p.45Chapter 2.5.1. --- Surgical Procedure - Ovariectomy (OVX) --- p.45Chapter 2.5.2. --- Surgical Procedure - Raloxifene Tubing Insertion --- p.45Chapter 2.5.3. --- High-Cholesterol Food Preparation --- p.45Chapter 2.5.4. --- "Body Weight, Food Consumption and Uterine Weight" --- p.46Chapter 2.5.5. --- Measurement of Raloxifene Tubing Consumption --- p.46Chapter 2.5.6. --- Serum Lipid and Lipoprotein Determinations --- p.46Chapter 2.5.7. --- Effect of Chronic Raloxifene on Artery Reactivity --- p.46Chapter 2.6. --- Solutions and Drugs --- p.49Chapter 2.6.1. --- "Drugs, Chemicals and Enzymes" --- p.49Chapter 2.6.2. --- Solutions --- p.51Chapter 2.6.3. --- Diet Composition for Syrian Golden Hamsters --- p.51Chapter 2.7. --- Statistical Analysis --- p.52Chapter CHAPTER III - --- "Raloxifene Relaxes Rat Pulmonary Arteries and Veins: Roles of Gender, Endothelium, and Antagonism of Ca Influx"Chapter 3.1. --- Abstract --- p.53Chapter 3.2. --- Introduction --- p.54Chapter 3.3. --- Methods and Materials --- p.55Chapter 3.3.1. --- Blood Vessel Preparation --- p.55Chapter 3.3.2. --- Protocols --- p.55Chapter 3.3.3. --- Measurement of Vascular Smooth Muscle [Ca2+]i --- p.56Chapter 3.3.4. --- Drugs --- p.57Chapter 3.3.5. --- Data Analysis --- p.53Chapter 3.4. --- Results --- p.58Chapter 3.4.1. --- Effects of Raloxifene on Pulmonary Arteries --- p.53Chapter 3.4.2. --- Effect of Raloxifene on CaCl2-induced Constrictionin Pulmonary Arteries --- p.59Chapter 3.4.3. --- Effects of Raloxifene on Pulmonary Veins --- p.59Chapter 3.4.4. --- Effect of Raloxifene on CaCl2-stimulated Increases in [Ca2+]i in Pulmonary Arteries --- p.60Chapter 3.5. --- Discussion --- p.67Chapter 3.6. --- Conclusion --- p.69Chapter CHAPTER IV - --- Raloxifene Modulates Pulmonary Vascular Reactivity in Spontaneously Hypertensive RatsChapter 4.1. --- Abstract --- p.70Chapter 4.2. --- Introduction --- p.71Chapter 4.3. --- Methods and Materials --- p.72Chapter 4.3.1. --- Raloxifene Treatment --- p.72Chapter 4.3.2. --- Blood Vessel Preparation --- p.72Chapter 4.3.3. --- Protocols --- p.73Chapter 4.3.4. --- Chemicals and Drugs --- p.73Chapter 4.3.5. --- Data Analysis --- p.74Chapter 4.4. --- Results --- p.74Chapter 4.4.1. --- Blood Pressure --- p.74Chapter 4.4.2. --- Vasocontraction in Spontaneously Hypertensive Rats --- p.75Chapter 4.4.3. --- Vasorelaxation in Spontaneously Hypertensive Rats --- p.75Chapter 4.4.4. --- Vasocontraction in Wistar-Kyoto rats --- p.76Chapter 4.4.5. --- Vasorelaxation in Wistar-Kyoto rats --- p.77Chapter 4.4.6. --- Comparison of contraction between WKY and SHR rats --- p.78Chapter 4.4.7. --- Comparison of relaxation between WKY and SHR rats --- p.78Chapter 4.5. --- Discussion --- p.93Chapter 4.6. --- Conclusion --- p.96Chapter CHAPTER V - --- Effects of Therapeutic Concentrations of Raloxifene in Pressurized Rat Small Mesenteric ArteryChapter 5.1. --- Abstract --- p.98Chapter 5.2. --- Introduction --- p.99Chapter 5.3. --- Methods and Materials --- p.101Chapter 5.3.1. --- Blood Vessel Preparation --- p.101Chapter 5.3.2. --- Experimental Protocols --- p.102Chapter 5.3.2.1. --- Myogenic Tone Development --- p.102Chapter 5.3.2.2. --- Effects of Raloxifene and 17β-EstradioI on Myogenic Constriction --- p.102Chapter 5.3.2.3. --- Effects of Pharmacological Inhibitors on Raloxifene- or 17β-Estradiol-induced Myogenic Constriction --- p.103Chapter 5.3.3. --- Drugs and Solutions --- p.103Chapter 5.3.4. --- Expression of Results and Statistical Analysis --- p.104Chapter 5.4. --- Results --- p.104Chapter 5.4.1. --- Effects of Raloxifene and 17β-Estradiol on Rat Resistance Mesenteric Arteries1 --- p.104Chapter 5.4.2. --- Effects of Inhibitors of NOS --- p.105Chapter 5.4.3. --- Effect of CTX plus Apamin --- p.106Chapter 5.4.4. --- "Effect of ICI 182,780" --- p.106Chapter 5.4.5. --- "Effects of Wortmannin, LY 294002 and Cycloheximide" --- p.106Chapter 5.5. --- Discussion --- p.122Chapter 5.6. --- Conclusion --- p.125Chapter CHAPTER VI - --- Effects of Chronic Raloxifene Treatment on Vascular Reactivity in Pressurized Septal Coronary Arteries from Hamsters Fed with High-Cholesterol DietChapter 6.1. --- Abstract --- p.127Chapter 6.2. --- Introduction --- p.128Chapter 6.3. --- Methods and Materials --- p.129Chapter 6.3.1. --- Preparatory Work --- p.129Chapter 6.3.1.1. --- Animals and Diets --- p.129Chapter 6.3.1.2. --- Preparation of High-Cholesterol (HC) Food --- p.129Chapter 6.3.1.3. --- Surgical Procedure - Ovariectomy (OVX) --- p.129Chapter 6.3.1.4. --- Surgical Procedure - Raloxifene Tubing Insertion --- p.130Chapter 6.3.1.5. --- Blood Vessel Preparation --- p.130Chapter 6.3.1.6. --- "Body Weight, Food Consumption and Uterine Weight" --- p.131Chapter 6.3.1.7. --- Measurement of Raloxifene Tubing Consumption --- p.131Chapter 6.3.1.8. --- Serum Lipid and Lipoprotein Determinations --- p.132Chapter 6.3.2. --- Experimental Protocols --- p.132Chapter 6.3.2.1. --- Development of Myogenic Tone --- p.132Chapter 6.3.2.2. --- Pressure-Diameter Relationships --- p.132Chapter 6.3.2.3. --- The Effect of Acetylcholine --- p.133Chapter 6.3.2.4. --- The Effect of U46619 --- p.133Chapter 6.3.2.5. --- The Effect of L-NAME --- p.133Chapter 6.3.3. --- Drugs and Solutions --- p.133Chapter 6.3.4. --- Expression of Results and Statistical Analysis --- p.134Chapter 6.4. --- Results --- p.135Chapter 6.4.1. --- Effects on Myogenic Response --- p.135Chapter 6.4.2. --- "Effects of Acetylcholine, U46619 and L-NAME" --- p.135Chapter 6.4.2.1. --- Comparison between OHHCD and OvxOHHCD --- p.135Chapter 6.4.2.2. --- Comparison between OvxOHHCD and OvxOHHCDRf --- p.135Chapter 6.4.2.3. --- Comparison between OHHCDRf and OvxOHHCDRf --- p.136Chapter 6.4.2.4. --- Comparison between OHHCD and OHHCDRf --- p.136Chapter 6.5. --- Discussion --- p.155Chapter 6.6. --- Conclusion --- p.156References --- p.157Publications --- p.17