Application of ion channel modulators in the reduction of triamcinolone cytotoxicity.

Zheng Tingting.Thesis (M.Phil.)--Chinese University of Hong Kong, 2004.Includes bibliographical references (leaves 98-124).Abstracts in English and Chinese.Abstract --- p.iAcknowledgements --- p.ivTable of Contents --- p.viList of Tables --- p.viiiList of Figures --- p.ixAbbreviations --- p.xiChapter Chapter 1 --- Introduction --- p.1Chapter 1.1 --- Triamcinolone acetonide(TA) --- p.1Chapter 1.1.1 --- Application in ophthalmology --- p.1Chapter 1.1.2 --- Mechanism of anti-inflammatory effect --- p.2Chapter 1.1.3 --- Side effects of TA --- p.4Chapter 1.1.4 --- Toxicity of TA --- p.5Chapter 1.2 --- Retinal pigment epithelial (RPE) cell --- p.6Chapter 1.3 --- Mechanism of cell death --- p.8Chapter 1.3.1 --- Apoptosis --- p.9Chapter 1.3.2 --- caspase --- p.11Chapter 1.3.3 --- Mitogen-activated protein kinase (MAPK) --- p.13Chapter 1.3.4 --- Activator Protein-1 (AP-1) --- p.15Chapter 1.4 --- Potassium channel (K+）） --- p.15Chapter 1.4.1 --- Molecular structure of KAtp channel --- p.16Chapter 1.4.2 --- Regulation of Katp channel --- p.17Chapter 1.4.3 --- Pinacidil --- p.18Chapter 1.5 --- Calcium channel --- p.20Chapter 1.5.1 --- VDCCs and subtypes --- p.21Chapter 1.5.2 --- Calcium channel blocker --- p.22Chapter 1.5.3 --- Verapamil --- p.23Chapter 1.6 --- Study objectives --- p.25Chapter Chapter 2 --- Methodology --- p.37Chapter 2.1 --- Cell biology --- p.37Chapter 2.1.1 --- Materials --- p.37Chapter 2.1.1.1 --- Culture related material --- p.37Chapter 2.1.1.2 --- Drugs --- p.37Chapter 2.1.1.3 --- Cell line and instrument --- p.37Chapter 2.1.2 --- Preparations --- p.38Chapter 2.1.2.1 --- Working medium --- p.38Chapter 2.1.2.2 --- Drugs --- p.38Chapter 2.1.2.3 --- MTT solution --- p.39Chapter 2.1.3 --- Cell culture and treatment process --- p.40Chapter 2.1.3.1 --- Seed cell --- p.40Chapter 2.1.3.2 --- Treatment --- p.40Chapter 2.1.4 --- MTT-Cell Proliferation Assay --- p.41Chapter 2.2 --- Molecular biology --- p.42Chapter 2.2.1 --- Materials --- p.42Chapter 2.2.1.1 --- "Chemicals, reagents, and kits" --- p.42Chapter 2.2.1.2 --- Solutions and Buffers --- p.42Chapter 2.2.1.3 --- Primers and Enzymes --- p.43Chapter 2.2.1.4 --- Equipment --- p.43Chapter 2.2.1.5 --- Software --- p.43Chapter 2.2.2 --- Reverse transcription 226}0ؤ Polymerase Chain Reaction (RT-PCR) --- p.44Chapter 2.2.2.1 --- Cell collection and RNA Isolation --- p.44Chapter 2.2.2.2 --- Reverse Transcription (RT) --- p.45Chapter 2.2.2.3 --- PCR Reaction --- p.46Chapter 2.3 --- Immunocytochemistry --- p.48Chapter 2.3.1 --- Materials and instrumentation --- p.49Chapter 2.3.1.1 --- Antibodies and Equipment --- p.49Chapter 2.3.1.2 --- Chemicals and other useful items --- p.49Chapter 2.3.2 --- Preparations --- p.50Chapter 2.3.2.1 --- Preparation of coverslips --- p.50Chapter 2.3.2.2 --- Prepations of solutions --- p.50Chapter 2.3.3 --- Procedures --- p.51Chapter 2.4 --- Expression of results and statistics --- p.52Chapter Chapter 3 --- Results --- p.57Chapter 3.1 --- Effects of TA on RPE cell culture --- p.57Chapter 3.1.1 --- Cell morphology --- p.57Chapter 3.1.2 --- MTT assay --- p.57Chapter 3.1.3 --- Gene expressions --- p.57Chapter 3.2 --- Effects of PIN/VP on TA treated RPE cells --- p.58Chapter 3.2.1 --- MTT assay --- p.58Chapter 3.2.2 --- Gene expression --- p.59Chapter 3.2.2.1 --- Expression of housekeeping gene --- p.59Chapter 3.2.2.2 --- Expression of apoptosis-related gene --- p.59Chapter 3.2.2.3 --- Expression of early-response genes --- p.60Chapter 3.2.3 --- Immunofluorescence --- p.61Chapter Chapter 4 --- Discussion --- p.86Chapter Chapter 5 --- References --- p.9