Pharmacological characterization of prehispanolone, a PAF receptor antagonist.

by Chu, Pui-ya.Thesis (M.Phil.)--Chinese University of Hong Kong.Bibliography: leaves 115-126.ACKNOWLEDGEMENTLIST OF ABBREVIATIONSABSTRACT --- p.1Chapter CHAPTER 1 --- INTRODUCTIONChapter 1.1 --- PLATELET-ACTIVATING FACTOR (PAF) --- p.4Chapter 1.1.1 --- Metabolism of PAF --- p.4Chapter 1.1.1.1 --- Biosynthesis of PAF --- p.4Chapter 1.1.1.2 --- Degradation of PAF --- p.9Chapter 1.1.2 --- Systemic effects of PAF --- p.12Chapter 1.1.3 --- Structure - activity relationship of PAF --- p.16Chapter 1.2 --- PAF RECEPTORS --- p.19Chapter 1.2.1 --- PAF receptor on platelets --- p.19Chapter 1.2.2 --- PAF receptor on leukocytes --- p.20Chapter 1.2.3 --- PAF receptor on macrophages --- p.20Chapter 1.2.4 --- Antagonists of PAF --- p.20Chapter 1.2.4.1 --- Nonspecific inhibitors of PAF --- p.21Chapter 1.2.4.2 --- Specific antagonists of PAF --- p.21Chapter 1.3 --- SECOND MESSENGER SYSTEMS OF PAF --- p.22Chapter 1.3.1 --- Adenylate cyclase - cyclic adenosine mono- phosphate system --- p.26Chapter 1.3.2 --- Phospholipase C - phosphatidylinositol system --- p.28Chapter 1.3.3 --- Intracellular calcium --- p.29Chapter 1.3.4 --- The role of prostaglandins and leukotrienes --- p.30Chapter 1.3.5 --- Protein kinase C --- p.30Chapter 1.3.5.1 --- Dual action of PKC --- p.31Chapter 1.3.5.2 --- Down regulation of PKC --- p.33Chapter 1.4 --- THE FUNCTION OF MACROPHAGES --- p.33Chapter 1.4.1 --- Phagocytosis --- p.34Chapter 1.4.2 --- Antigen presentation --- p.34Chapter 1.4.3 --- Release of cytokines --- p.34Chapter 1.4.4 --- Respiratory burst --- p.35Chapter CHAPTER 2 --- METHODSChapter 2.1 --- PREPARATION OF PERITONEAL MACROPHAGES --- p.37Chapter 2.1.1 --- Preparation of reagents --- p.37Chapter 2.1.2 --- Preparation of peritoneal macrophages --- p.37Chapter 2.2 --- RADIOLIGAND BINDING ASSAY --- p.38Chapter 2.2.1 --- Reagents --- p.38Chapter 2.2.2 --- [3H]-PAF binding to TG-PEC --- p.39Chapter 2.2.2.1 --- Preparation of thioglycollate-elicited peritoneal macrophages --- p.39Chapter 2.2.2.2 --- Preparation of working solutions --- p.39Chapter 2.2.2.3 --- Assay of [3H]-PAF binding --- p.40Chapter 2.2.3 --- [3H]-PAF binding to resident PEC --- p.40Chapter 2.2.3.1 --- Preparation of resident peritoneal mcrophages --- p.41Chapter 2.2.3.2 --- Preparation of working solutions --- p.41Chapter 2.2.3.3 --- Assay of [3H]-PAF binding --- p.41Chapter 2.2.4 --- Preparation of drugs --- p.41Chapter 2.2.5 --- Data analysis --- p.42Chapter 2.3 --- MEASUREMENT OF INOSITOL PHOSPHATES --- p.42Chapter 2.3.1 --- Preparation of reagents --- p.42Chapter 2.3.2 --- Dowex column preparation --- p.43Chapter 2.3.3 --- Cell plating in 24 - well plastic trays --- p.44Chapter 2.3.4 --- Determination of total inositol phosphates --- p.44Chapter 2.3.5 --- Column separation --- p.45Chapter 2.3.6 --- Determination of inositol trisphosphate (IP3) --- p.46Chapter 2.4 --- MEASUREMENT OF INOSITOL PHOSPHATES ACCUMULATION AFTER PROLONGED PMA PRETREATMENT --- p.46Chapter 2.4.1 --- Preparation of phorbol-12-myristate-13-acetate (PMA) solutions --- p.47Chapter 2.4.2 --- Preparation of cells with PMA --- p.47Chapter 2.5 --- DETERMINATION OF SUPEROXIDE ANION PRODUCTION --- p.47Chapter 2.5.1 --- Preparation of drugs --- p.47Chapter 2.5.2 --- Assay of superoxide anion production --- p.48Chapter 2.5.3 --- Data analysis --- p.48Chapter 2.6 --- STATISTICAL METHOD --- p.48Chapter CHAPTER 3 --- RESULTSChapter 3.1 --- [3H]-PAF BINDING TO TG-PEC --- p.50Chapter 3.1.1 --- General properties --- p.50Chapter 3.1.2 --- Stereospecific of PAF receptor --- p.57Chapter 3.1.3 --- Comparison of [3H]-PAF binding to TG-PEC and TG-PMΦ from Balb/c mice --- p.57Chapter 3.1.4 --- Inhibition of specific binding of [3H]-PAF to murine and guinea pig TG-PEC by PAF --- p.57Chapter 3.1.5 --- Effects of PAF antagonists on the binding of [3H]- PAF to TG-PEC from Balb/c mice and guinea pigs --- p.60Chapter 3.1.6 --- Scatchard analysis of [3H]-PAF binding to murine and guinea pig TG-PMΦ --- p.65Chapter 3.2 --- SUPEROXIDE ANION PRODUCTION IN PERITONEAL MACROPHAGES --- p.69Chapter 3.2.1 --- Superoxide anion production induced by PAFin murine and guinea pig TG-PMΦ --- p.69Chapter 3.2.2 --- Effect of PMA on superoxide anion production in murine and guinea pig TG-PMΦ --- p.69Chapter 3.2.3 --- Effect of calcium ionophore on superoxide anion production in murine and guinea pig TG-PMΦ --- p.75Chapter 3.2.4 --- Effect of PAF antagonists on PAF - induced superoxide anion production in guinea pig TG-PMΦ --- p.75Chapter 3.3 --- PHOSPHOLIPASE C - PHOSPHATIDYLINOSITOL SYSTEM IN PERITONEAL MACROPHAGES --- p.79Chapter 3.3.1 --- Effect of PAF on the accumulation of inositol phosphates in the absence of LiCl --- p.79Chapter 3.3.2 --- Effect of PAF on the accumulation of inositol phosphates in the presence of LiCl --- p.79Chapter 3.3.2.1 --- Time course of [3H]-inositol phosphates accumulation induced by PAF in TG-PMΦ --- p.86Chapter 3.3.2.2 --- Effect of PAF on the accumulation of [3H]-IPs in TG- PMΦ from Balb/c mice and guinea pigs --- p.86Chapter 3.3.2.3 --- Effect of PAF antagonists on PAF-induced [3H]-IPs accumulation in TG-PMΦ from Balb/c mice and guinea pigs --- p.86Chapter 3.3.2.4 --- Effect of PMA on PAF-induced [3H]-IPs formation in TG-PMΦ from Balb/c mice and guinea pigs --- p.90Chapter 3.3.2.5 --- Effect of prolonged PMA pretreatment on PAF and PMA-induced [3H]-IPs accumulation in murine and guinea pig TG-PMΦ --- p.90Chapter 3.4 --- STUDIES OF RESIDENT PEC FROM Balb/c MICE --- p.96Chapter 3.4.1 --- Binding of 2nM [3H]-PAF to Balb/c mice resident PEC --- p.96Chapter 3.4.2 --- PAF-induced [3H]-IPs accumulation in murine resident PMΦ --- p.100Chapter 3.4.3 --- Binding of 0.2 nM [3H]-PAF to Balb/c mice resident PEC --- p.100Chapter 3.4.4 --- Superoxide anion production induced by PAF and PMA in murine resident PMΦ --- p.104Chapter CHAPTER 4 --- DISCUSSIONSChapter 4.1 --- PAF RECEPTOR ON PERITONEAL MACROPHAGES --- p.125Chapter 4.1.1 --- [3H]-PAF binding to peritoneal macrophages --- p.105Chapter 4.1.2 --- Expression of PAF receptor on Balb/c mice peritoneal macrophages --- p.107Chapter 4.2 --- SUPEROXIDE ANION PRODUCTION IN PERITONEAL MACROPHAGES --- p.108Chapter 4.3 --- PAF RECEPTOR AND POLYPHOSPHATIDYLINOSITOL SYSTEM IN PERITONEAL MACROPHAGES --- p.109Chapter CHAPTER 5 --- CONCLUSIONS --- p.112REFERENCES --- p.11