The effects of phosphodiesterase inhibitors on rat mast cells.

• Other Authors: Kam, Man Fai Afia.
• Format: Others
• Language: English; Chinese
• Published: 2005
• Subjects: Mast cells > Physiology; Phosphodiesterases > Inhibitors; Histamine Release > drug effects; Mast Cells > drug effects; Phosphodiesterase Inhibitors > therapeutic use
• Online Access: http://library.cuhk.edu.hk/record=b5892476; http://repository.lib.cuhk.edu.hk/en/item/cuhk-325327

Kam Man Fai Afia. === Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. === Includes bibliographical references (leaves [195]-224). === Abstracts in English and Chinese. === Abstract --- p.i === Acknowledgement --- p.v === Publications --- p.vi === Abbreviations --- p.vii === Chapter 1. --- Introduction --- p.1 === Chapter 1.1 --- The Mast Cell --- p.2 === Chapter 1.1.1 --- Historical Perspective --- p.2 === Chapter 1.1.2 --- Mast Cell Origin and Development --- p.3 === Chapter 1.1.3 --- Mast Cell Heterogeneity --- p.5 === Chapter 1.1.3.1 --- Rodent Mast Cell Heterogeneity --- p.5 === Chapter 1.1.3.2 --- Human Mast Cell Heterogeneity --- p.7 === Chapter 1.1.4 --- Mast Cell Mediators --- p.10 === Chapter 1.1.4.1 --- Preformed Mediators --- p.11 === Chapter 1.1.4.2 --- Newly Synthesized Lipid Mediators --- p.14 === Chapter 1.1.4.3 --- Cytokines --- p.16 === Chapter 1.1.5 --- Mast Cell Activation --- p.17 === Chapter 1.1.5.1 --- Immunological Activation --- p.19 === Chapter 1.1.5.1.1 --- FcεIR Activation and Protein Tyrosine Phosphorylation --- p.19 === Chapter 1.1.5.1.2 --- Activation of Phospholipases --- p.20 === Chapter 1.1.5.1.3 --- The Role of Calcium --- p.22 === Chapter 1.1.5.1.3.1 --- Intracellular Calcium Mobilization --- p.23 === Chapter 1.1.5.1.3.2 --- Calcium Influx --- p.24 === Chapter 1.1.5.1.3.3 --- Mechanisms of Action of Calcium in Mast Cells --- p.28 === Chapter 1.1.5.1.4 --- The Role of G-proteins --- p.30 === Chapter 1.1.5.1.5. --- The Role of Cylic AMP --- p.33 === Chapter 1.1.5.1.2.1 --- Mechanisms of Action of Cyclic AMP in Mast Cells --- p.36 === Chapter 1.1.5.1.2.2 --- Implications for the Inhibitory Role of Cyclic AMP in Mast Cell Activation --- p.37 === Chapter 1.2 --- The Cyclic Nucleotide Phosphodiesterases --- p.39 === Chapter 1.2.1 --- Introduction --- p.39 === Chapter 1.2.2 --- Classification and Structure --- p.41 === Chapter 1.2.3 --- Distribution and Physiological Functions of the Different PDE Families --- p.45 === Chapter 1.2.4 --- Phosphodiesterase Inhibitors --- p.49 === Chapter 1.2.4.1 --- Non-selective PDE Inhibitors --- p.50 === Chapter 1.2.4.2 --- Selective PDE Inhibitors --- p.52 === Chapter 1.2.4.2.1 --- PDE1 and PDE2 Inhibitors --- p.52 === Chapter 1.2.4.2.2 --- PDE3 Inhibitors --- p.53 === Chapter 1.2.4.2.3 --- PDE4 Inhibitors --- p.54 === Chapter 1.2.4.2.4.1 --- PDE5 Inhibitors --- p.56 === Chapter 2. --- Materials and Methods --- p.59 === Chapter 2.1 --- Materials --- p.60 === Chapter 2.1.1 --- Drugs --- p.60 === Chapter 2.1.1.1 --- Phosphodiesterase Inhibitors --- p.60 === Chapter 2.1.1.2 --- Mast Cell Secretagogues --- p.61 === Chapter 2.1.2 --- Materials for Rat Peritoneal Mast Cell Experiments --- p.61 === Chapter 2.1.2.1 --- Materials for Rat Sensitization --- p.61 === Chapter 2.1.2.2 --- Materials for Buffers --- p.62 === Chapter 2.1.2.3 --- Materials for Histamine Assay --- p.62 === Chapter 2.1.2.4 --- Miscellaneous --- p.63 === Chapter 2.1.3 --- Materials for RBL-2H3 Cell Line Experiments --- p.63 === Chapter 2.1.3.1 --- Materials for Cell Culture --- p.63 === Chapter 2.1.3.2 --- Materials for Cell Sensitization and Enzyme Release --- p.64 === Chapter 2.1.3.3 --- Materials for β-Hexosaminidase Assay --- p.64 === Chapter 2.1.3.4 --- Miscellaneous --- p.64 === Chapter 2.2 --- Rat Peritoneal Mast Cell Experiments --- p.65 === Chapter 2.2.1 --- Preparation of Buffers --- p.65 === Chapter 2.2.2 --- Preparation of Stock Solutions --- p.66 === Chapter 2.2.2.1 --- Mast Cell Secretagogue Stock Solutions --- p.66 === Chapter 2.2.2.2 --- Phosphodiesterase Inhibitor Stock Solutions --- p.66 === Chapter 2.2.3 --- Animals and Cell Isolation --- p.71 === Chapter 2.2.3.1 --- Animals --- p.71 === Chapter 2.2.3.2 --- Sensitization of Animals --- p.71 === Chapter 2.2.3.3 --- Cell Isolation --- p.71 === Chapter 2.2.3.4 --- Cell Purification --- p.72 === Chapter 2.2.3.5 --- Determination of Cell Number and Viability --- p.73 === Chapter 2.2.4 --- General Protocol for Histamine Release and Histamine Measurement --- p.75 === Chapter 2.2.4.1 --- Histamine Release --- p.75 === Chapter 2.2.4.2 --- Spectrofluorometric Determination of Histamine Content --- p.76 === Chapter 2.2.4.2.1 --- Manual Histamine Assay --- p.76 === Chapter 2.2.4.2.2 --- Automated Histamine Assay --- p.78 === Chapter 2.2.4.3 --- Calculation of Histamine Levels --- p.78 === Chapter 2.2.4.4 --- Presentation and Statistics --- p.79 === Chapter 2.3 --- RBL-2H3 Cell Line Experiments --- p.80 === Chapter 2.3.1 --- Preparation of Stock Solutions --- p.80 === Chapter 2.3.2 --- Preparation of Materials for Enzyme Release and Assay --- p.81 === Chapter 2.3.2.1 --- Cell Culture --- p.81 === Chapter 2.3.2.2 --- Preparation of Cells for β-Hexosaminidase Release Experiments --- p.82 === Chapter 2.3.2.3 --- β-Hexosaminidase Release --- p.82 === Chapter 2.3.2.4 --- β-Hexosaminidase Assay --- p.83 === Chapter 3. --- Effects of Phosphodiesterase Inhibitors on Mediator Release from Rat Mast Cells --- p.84 === Chapter 3.1 --- Introduction --- p.85 === Chapter 3.2 --- Materials and Methods --- p.87 === Chapter 3.2.1 --- Rat Peritoneal Mast Cells --- p.87 === Chapter 3.2.1.1 --- Experiments Employing Immunological Stimulus in RPMCs --- p.87 === Chapter 3.2.1.2 --- Experiments Employing Non-Immunological Stimuli in RPMCs --- p.88 === Chapter 3.2.2 --- Rat Basophilic Leukemia Cells --- p.88 === Chapter 3.3 --- Results --- p.89 === Chapter 3.3.1 --- Rat Peritoneal Mast Cells --- p.89 === Chapter 3.3.1.1 --- Immunologically Activated Rat Peritoneal Mast Cells --- p.89 === Chapter 3.3.1.1.1 --- Effects of Non-Selective PDE Inhibitors on Anti-IgE-Mediated Histamine Release from RPMCs --- p.89 === Chapter 3.3.1.1.2 --- Effects of Selective PDE1 and PDE2 Inhibitors on Anti-IgE- Mediated Histamine Release from RPMCs --- p.90 === Chapter 3.3.1.1.3 --- Effects of Selective PDE3 Inhibitors on Anti-IgE-Mediated Histamine Release from RPMCs --- p.90 === Chapter 3.3.1.1.4 --- Effects of Selective PDE4 Inhibitors on Anti-IgE-Mediated Histamine Release from RPMCs --- p.91 === Chapter 3.3.1.1.5 --- Effects of Selective PDE5 Inhibitors on Anti-IgE-Mediated Histamine Release from RPMCs --- p.91 === Chapter 3.3.1.2 --- Non-Immunologically Activated Rat Peritoneal Mast Cells --- p.92 === Chapter 3.3.1.2.1 --- Effects of Selective PDE Inhibitors on Compound 48/80- Mediated Histamine Release from RPMCs --- p.92 === Chapter 3.3.1.2.2 --- Effects of Selective PDE Inhibitors on Histamine Release from RPMCs Stimulated by Calcium Ionophores --- p.93 === Chapter 3.3.2 --- Rat Basophilic Leukemia Cells --- p.93 === Chapter 3.3.2.1 --- Effects of Non-Selective PDE Inhibitors on Antigen-Mediated β-Hexosaminidase Release from RBL-2H3 Cells --- p.93 === Chapter 3.3.2.2 --- Effects of Selective PDE Inhibitors on Antigen-Mediated β-Hexosaminidase Release from RBL-2H3 Cells --- p.94 === Chapter 3.4 --- Discussion --- p.95 === Chapter 3.4.1 --- Rat Peritoneal Mast Cells --- p.95 === Chapter 3.4.1.1 --- Immunologically Activated RPMCs --- p.95 === Chapter 3.4.1.2 --- Non-Immunologically Activated RPMCs --- p.99 === Chapter 3.4.2 --- Rat Basophilic Leukemia Cells --- p.103 === Chapter 4. --- Combined Effects of Selective Phosphodiesterase Inhibitors on Immunologically Induced Histamine from Rat Mast Cells --- p.143 === Chapter 4.1 --- Introduction --- p.144 === Chapter 4.2 --- Materials and Methods --- p.144 === Chapter 4.2.1 --- Simultaneous Addition of PDE3 and PDE4 Inhibitors --- p.145 === Chapter 4.2.2 --- Sequential Addition of PDE3 and PDE4 Inhibitors --- p.145 === Chapter 4.3 --- Results --- p.146 === Chapter 4.3.1 --- Effects of the Selective Inhibitors for PDE3 and PDE4 Alone: Calculation of the Expected Inhibition Curve --- p.146 === Chapter 4.3.2 --- Effects of the Simultaneous Addition of PDE3 and PDE4 Inhibitors on Anti-IgE-Mediated Histamine Release from RPMCs --- p.148 === Chapter 4.3.2.1 --- Rolipram and Siguazodan --- p.148 === Chapter 4.3.2.2 --- Ro 20-1724 and Siguazodan --- p.149 === Chapter 4.3.2.3 --- Rolipram and Quazinone --- p.149 === Chapter 4.3.2.4 --- Ro 20-1724 and Quazinone --- p.150 === Chapter 4.3.3 --- Effects of the Sequential Addition of PDE3 and PDE4 Inhibitors on Anti-IgE-Mediated Histamine Release from RPMCs --- p.150 === Chapter 4.3.3.1 --- Rolipram and Siguazodan --- p.150 === Chapter 4.3.3.2 --- Ro 20-1724 and Siguazodan --- p.151 === Chapter 4.3.3.3 --- Rolipram and Quazinone --- p.151 === Chapter 4.3.3.4 --- Ro 20-1724 and Quazinone --- p.152 === Chapter 4.4 --- Discussion --- p.153 === Chapter 5. --- Future Directions --- p.191 === Chapter 5.1 --- Future Directions --- p.192 === References --- p.195