Study on the vascular actions of sulfonylurea drugs.

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 --- IntroductionChapter 1.1. --- Sulfonylureas --- p.1Chapter 1.2. --- Biological action of sulfonylurea drugs --- p.2Chapter 1.2.1 --- Effects on pancreatic β cells --- p.5Chapter 1.2.2. --- Effects on cardiac myocytes --- p.7Chapter 1.2.3. --- Effects on smooth muscle cells --- p.11Chapter 1.2.4. --- Effects on endothelial cells --- p.14Chapter 1.3. --- Side effects and toxicity --- p.15Chapter 1.4. --- Objectives of the present study --- p.17Chapter Chapter 2 --- Methods and MarterialsChapter 2.1. --- Tissue and Cell Preparation --- p.20Chapter 2.1.1. --- Preparation of the isolated rat aorta and mesenteric artery --- p.20Chapter 2.1.2. --- Removal of the functional endothelium --- p.20Chapter 2.1.3. --- Cell culture --- p.21Chapter 2.1.3.1. --- Materials --- p.21Chapter 2.1.3.2. --- Aortic smooth muscle cells in primary culture --- p.21Chapter 2.1.3.3. --- Aortic endothelial cells in primary culture --- p.23Chapter 2.1.3.4. --- Cultured rat aortic smooth muscle cell line (A7r5) --- p.23Chapter 2.1.3.5. --- Cultured human umbilical vein endothelial cell line (ECV-304) --- p.24Chapter 2.1.3.6. --- Cell subculture --- p.24Chapter 2.1.3.7. --- Immunostaining of endothelial cells in primary culture --- p.24Chapter 2.2. --- Organ Bath Set-up --- p.25Chapter 2.3. --- Force Measurement --- p.28Chapter 2.3.1. --- Vascular action of glibenclamide --- p.28Chapter 2.3.1.1. --- Antagonistic effect of glibenclamide on relaxation induced by K+ channel openers --- p.28Chapter 2.3.1.2. --- Relaxant response of glibenclamide --- p.29Chapter 2.3.1.3. --- Role of endothelium-derived vasoactive factors in glibenclamide induced relaxation --- p.29Chapter 2.3.1.4. --- Effect of endothelial prostanoids in glibenclamide-induced relaxation --- p.30Chapter 2.3.1.5. --- Effects of putative K+ channel blockers on glibenclamide-induced relaxation --- p.30Chapter 2.3.1.6. --- Effect of glibenclamide on high K+- and CaCl2-induced contraction --- p.31Chapter 2.3.1.7. --- Effect of glibenclamide on prostaglandin F2α-induced contraction --- p.32Chapter 2.3.1.8. --- Effect of glibenclamide on protein kinase C-mediated contraction --- p.32Chapter 2.3.2. --- Vascular action of glipizide --- p.33Chapter 2.3.3. --- Vascular action of tolbutamide --- p.33Chapter 2.3.3.1. --- Contractile response of tolbutamide --- p.33Chapter 2.3.3.2. --- Effects of inhibitors of endothelium-derived factors --- p.33Chapter 2.3.3.3. --- Effects of inhibitors of Ca2+ influx --- p.34Chapter 2.3.3.4. --- Effect of protein kinase C inhibitor --- p.34Chapter 2.3.3.5. --- Effects of neural factors --- p.34Chapter 2.4. --- Cyclic GMP measurement --- p.35Chapter 2.4.1. --- Material --- p.35Chapter 2.4.2. --- Methods --- p.35Chapter 2.4.2.1. --- Tissue preparation --- p.35Chapter 2.4.2.2. --- Plasma and tissue according to protocols provided by Amersham --- p.35Chapter 2.4.2.3. --- Cyclic GMP content measurement --- p.36Chapter 2.4.2.4. --- Protein content measurement --- p.39Chapter 2.4.2.5. --- Cyclic GMP measurement protocol --- p.40Chapter 2.5. --- Ca2+ measurement --- p.40Chapter 2.5.1. --- Materials --- p.40Chapter 2.5.1.1. --- PTI RatioMaster Fluorescence System --- p.40Chapter 2.5.1.2. --- Confocal Imaging System --- p.42Chapter 2.5.2. --- Method --- p.42Chapter 2.5.3. --- Protocols for Ca2+ measurement --- p.45Chapter 2.5.3.1. --- Effect of glibenclamide in endothelial cells --- p.45Chapter 2.5.3.2. --- Effect of glibenclamide in vascular smooth muscle cells --- p.45Chapter 2.5.3.3. --- Effect of tolbutamide in vascular smooth muscle cells --- p.46Chapter 2.6. --- Cell proliferation --- p.45Chapter 2.6.1. --- Materials --- p.45Chapter 2.6.2. --- Method --- p.46Chapter 2.6.3. --- Protocols for cell proliferation --- p.47Chapter 2.6.3.1. --- Effect of glibenclamide on endothelial cell proliferation --- p.47Chapter 2.6.3.2. --- Effect of glibenclamide on aortic smooth muscle cell proliferation --- p.47Chapter 2.7. --- Chemicals and solutions --- p.48Chapter 2.8. --- Statistical analysis --- p.50Chapter Chapter 3 --- ResultsChapter 3.1. --- Glibenclamide --- p.51Chapter 3.1.1. --- Effect of glibenclamide on the K+ channel activity --- p.51Chapter 3.1.2. --- Relaxant response of glibenclamide --- p.55Chapter 3.1.3. --- Effects of inhibitors of nitric oxide activity on glibenclamide- induced relaxation --- p.57Chapter 3.1.4. --- Role of endothelial relaxing prostanoids in glibenclamide-induced relaxation --- p.69Chapter 3.1.5. --- Effect of putative K+ channel blockers on glibenclamide-induced relaxation --- p.73Chapter 3.1.6. --- Effect of glibenclamide on high K+-induced arterial contraction --- p.75Chapter 3.1.7. --- Effect of glibenclamide on protein kinase C-mediated contraction --- p.83Chapter 3.1.8. --- Effect of glibenclamide on prostaglandin F2α-induced contraction --- p.83Chapter 3.2 --- Glipizide --- p.85Chapter 3.2.1. --- Relaxant response of glipizide --- p.85Chapter 3.3. --- Tolbutamide --- p.91Chapter 3.3.1. --- Contractile response to tolbutamide --- p.91Chapter 3.3.2. --- Effects of endothelium-derived factors --- p.94Chapter 3.3.3. --- Effects of inhibitors of Ca2+ influx on tolbutamide-induced contraction --- p.98Chapter 3.3.4. --- "Effects of forskolin, sodium nitroprusside, staurosporine on tolbutamide-induced contraction" --- p.102Chapter 3.3.5. --- Effect of neural factors --- p.106Chapter 3.4. --- Effect of glibenclamide on cGMP levels --- p.112Chapter 3.5. --- Effect of glibenclamide on intracellular[Ca2+ ] in cultured endothelial cells --- p.112Chapter 3.6. --- Effect of glibenclamide on intracellular [Ca2+] in cultured aortic smooth muscle cells --- p.115Chapter 3.7. --- Effect of tolbutamide on intracellular [Ca2+] in cultured aortic smooth muscle cells --- p.121Chapter 3.8. --- Effect of glibenclamide on proliferation of cultured endothelial cells --- p.121Chapter 3.9. --- Effect of glibenclamide on proliferation of cultured aortic smooth muscle cells --- p.123Chapter Chapter 4 --- DiscussionChapter 4.1. --- Effect of glibenclamide --- p.133Chapter 4.2. --- Effect of glipizide --- p.143Chapter 4.3. --- Effect of tolbutamide --- p.144Chapter 4.4. --- Conclusion --- p.152References --- p.155Publications --- p.16