Wai Kei Chan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 155-164). / Abstracts in English and Chinese. / Chapter Chapter 1 --- Introduction / Chapter 1.1. --- Sulfonylureas --- p.1 / Chapter 1.2. --- Biological action of sulfonylurea drugs --- p.2 / Chapter 1.2.1 --- Effects on pancreatic β cells --- p.5 / Chapter 1.2.2. --- Effects on cardiac myocytes --- p.7 / Chapter 1.2.3. --- Effects on smooth muscle cells --- p.11 / Chapter 1.2.4. --- Effects on endothelial cells --- p.14 / Chapter 1.3. --- Side effects and toxicity --- p.15 / Chapter 1.4. --- Objectives of the present study --- p.17 / Chapter Chapter 2 --- Methods and Marterials / Chapter 2.1. --- Tissue and Cell Preparation --- p.20 / Chapter 2.1.1. --- Preparation of the isolated rat aorta and mesenteric artery --- p.20 / Chapter 2.1.2. --- Removal of the functional endothelium --- p.20 / Chapter 2.1.3. --- Cell culture --- p.21 / Chapter 2.1.3.1. --- Materials --- p.21 / Chapter 2.1.3.2. --- Aortic smooth muscle cells in primary culture --- p.21 / Chapter 2.1.3.3. --- Aortic endothelial cells in primary culture --- p.23 / Chapter 2.1.3.4. --- Cultured rat aortic smooth muscle cell line (A7r5) --- p.23 / Chapter 2.1.3.5. --- Cultured human umbilical vein endothelial cell line (ECV-304) --- p.24 / Chapter 2.1.3.6. --- Cell subculture --- p.24 / Chapter 2.1.3.7. --- Immunostaining of endothelial cells in primary culture --- p.24 / Chapter 2.2. --- Organ Bath Set-up --- p.25 / Chapter 2.3. --- Force Measurement --- p.28 / Chapter 2.3.1. --- Vascular action of glibenclamide --- p.28 / Chapter 2.3.1.1. --- Antagonistic effect of glibenclamide on relaxation induced by K+ channel openers --- p.28 / Chapter 2.3.1.2. --- Relaxant response of glibenclamide --- p.29 / Chapter 2.3.1.3. --- Role of endothelium-derived vasoactive factors in glibenclamide induced relaxation --- p.29 / Chapter 2.3.1.4. --- Effect of endothelial prostanoids in glibenclamide-induced relaxation --- p.30 / Chapter 2.3.1.5. --- Effects of putative K+ channel blockers on glibenclamide-induced relaxation --- p.30 / Chapter 2.3.1.6. --- Effect of glibenclamide on high K+- and CaCl2-induced contraction --- p.31 / Chapter 2.3.1.7. --- Effect of glibenclamide on prostaglandin F2α-induced contraction --- p.32 / Chapter 2.3.1.8. --- Effect of glibenclamide on protein kinase C-mediated contraction --- p.32 / Chapter 2.3.2. --- Vascular action of glipizide --- p.33 / Chapter 2.3.3. --- Vascular action of tolbutamide --- p.33 / Chapter 2.3.3.1. --- Contractile response of tolbutamide --- p.33 / Chapter 2.3.3.2. --- Effects of inhibitors of endothelium-derived factors --- p.33 / Chapter 2.3.3.3. --- Effects of inhibitors of Ca2+ influx --- p.34 / Chapter 2.3.3.4. --- Effect of protein kinase C inhibitor --- p.34 / Chapter 2.3.3.5. --- Effects of neural factors --- p.34 / Chapter 2.4. --- Cyclic GMP measurement --- p.35 / Chapter 2.4.1. --- Material --- p.35 / Chapter 2.4.2. --- Methods --- p.35 / Chapter 2.4.2.1. --- Tissue preparation --- p.35 / Chapter 2.4.2.2. --- Plasma and tissue according to protocols provided by Amersham --- p.35 / Chapter 2.4.2.3. --- Cyclic GMP content measurement --- p.36 / Chapter 2.4.2.4. --- Protein content measurement --- p.39 / Chapter 2.4.2.5. --- Cyclic GMP measurement protocol --- p.40 / Chapter 2.5. --- Ca2+ measurement --- p.40 / Chapter 2.5.1. --- Materials --- p.40 / Chapter 2.5.1.1. --- PTI RatioMaster Fluorescence System --- p.40 / Chapter 2.5.1.2. --- Confocal Imaging System --- p.42 / Chapter 2.5.2. --- Method --- p.42 / Chapter 2.5.3. --- Protocols for Ca2+ measurement --- p.45 / Chapter 2.5.3.1. --- Effect of glibenclamide in endothelial cells --- p.45 / Chapter 2.5.3.2. --- Effect of glibenclamide in vascular smooth muscle cells --- p.45 / Chapter 2.5.3.3. --- Effect of tolbutamide in vascular smooth muscle cells --- p.46 / Chapter 2.6. --- Cell proliferation --- p.45 / Chapter 2.6.1. --- Materials --- p.45 / Chapter 2.6.2. --- Method --- p.46 / Chapter 2.6.3. --- Protocols for cell proliferation --- p.47 / Chapter 2.6.3.1. --- Effect of glibenclamide on endothelial cell proliferation --- p.47 / Chapter 2.6.3.2. --- Effect of glibenclamide on aortic smooth muscle cell proliferation --- p.47 / Chapter 2.7. --- Chemicals and solutions --- p.48 / Chapter 2.8. --- Statistical analysis --- p.50 / Chapter Chapter 3 --- Results / Chapter 3.1. --- Glibenclamide --- p.51 / Chapter 3.1.1. --- Effect of glibenclamide on the K+ channel activity --- p.51 / Chapter 3.1.2. --- Relaxant response of glibenclamide --- p.55 / Chapter 3.1.3. --- Effects of inhibitors of nitric oxide activity on glibenclamide- induced relaxation --- p.57 / Chapter 3.1.4. --- Role of endothelial relaxing prostanoids in glibenclamide-induced relaxation --- p.69 / Chapter 3.1.5. --- Effect of putative K+ channel blockers on glibenclamide-induced relaxation --- p.73 / Chapter 3.1.6. --- Effect of glibenclamide on high K+-induced arterial contraction --- p.75 / Chapter 3.1.7. --- Effect of glibenclamide on protein kinase C-mediated contraction --- p.83 / Chapter 3.1.8. --- Effect of glibenclamide on prostaglandin F2α-induced contraction --- p.83 / Chapter 3.2 --- Glipizide --- p.85 / Chapter 3.2.1. --- Relaxant response of glipizide --- p.85 / Chapter 3.3. --- Tolbutamide --- p.91 / Chapter 3.3.1. --- Contractile response to tolbutamide --- p.91 / Chapter 3.3.2. --- Effects of endothelium-derived factors --- p.94 / Chapter 3.3.3. --- Effects of inhibitors of Ca2+ influx on tolbutamide-induced contraction --- p.98 / Chapter 3.3.4. --- "Effects of forskolin, sodium nitroprusside, staurosporine on tolbutamide-induced contraction" --- p.102 / Chapter 3.3.5. --- Effect of neural factors --- p.106 / Chapter 3.4. --- Effect of glibenclamide on cGMP levels --- p.112 / Chapter 3.5. --- Effect of glibenclamide on intracellular[Ca2+ ] in cultured endothelial cells --- p.112 / Chapter 3.6. --- Effect of glibenclamide on intracellular [Ca2+] in cultured aortic smooth muscle cells --- p.115 / Chapter 3.7. --- Effect of tolbutamide on intracellular [Ca2+] in cultured aortic smooth muscle cells --- p.121 / Chapter 3.8. --- Effect of glibenclamide on proliferation of cultured endothelial cells --- p.121 / Chapter 3.9. --- Effect of glibenclamide on proliferation of cultured aortic smooth muscle cells --- p.123 / Chapter Chapter 4 --- Discussion / Chapter 4.1. --- Effect of glibenclamide --- p.133 / Chapter 4.2. --- Effect of glipizide --- p.143 / Chapter 4.3. --- Effect of tolbutamide --- p.144 / Chapter 4.4. --- Conclusion --- p.152 / References --- p.155 / Publications --- p.163