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Bio-pharmacological study for the action of bioactive molecules which control the reciprocal relationship between mast cells and other inflammatory cells

控制肥大细胞与其他炎症细胞之间相互关系的生物活性分子作用的生物药理学研究

基本信息

批准号：
02404079
负责人：
ICHIKAWA Atsushi
金额：
$ 14.91万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (A)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1992
项目状态：
已结题

项目摘要

We have investigated the basic problems about the action mechanisms of bioactive molecules which control the reciprocal relation between mast cells and other inflammatory cells, focused on following three points. 1. Studies on histamine synthesis in mast cells. (1) Isolation of L-histidine decarboxylase from mouse mastocytoma P-815 cells was performed, and is cDNA-derived amino acid sequence was determined. (2) Synergistic effects of de novo synthesis of L-histidine decarboxylase and its mRNA was found in cultured mast cells treated with two different stimulants; dexamethasone and 12-O-tetradecanoylphorbol 13-acetate, or dibutyryl cAMP and calcium ionophore A23187. (3) Possible post-translational processing of L-histidine decarboxylase was suspected. 2. Studies on cellular interaction between mast cells and endothelial cells or platelets. (1) PGD_2, is metabolite DELTA^<12>-PGJ_2, from mast cells inhibited cell growth of, but induced 31kDa protein, which was hemeoxygenase, in endothelial cells. (2) PGI_2 from endothelial cells stimulated cAMP synthesis via Gs-coupled adenylate cyclase. (3) Differential regulation of thrombin- or ATP-induced mobilization of intracellular Ca_<2+> by PGI_2 receptor in mastocytoma cells. (4) PGI_2 receptors in mastocytoma cells and platelets were identified by use of irreversible specific photoaffinity probe. 3. Studies on structure and function of prostaglandin E receptor subtypes. and on their mRNA expression in various tissues. (1) Cloning and expression of cDNAs for mouse prostaglandin E receptors, EP_3 subtype and EP_2 subtypes. (2) Functional cDNA for two isoforms of PGE receptor EP_3 subtype with different C-terminal domains which were derived from alternative splicing were obtained. Two forms determined ligand-binding affinity and sensitivity of agonist-induced desensitization.

本文对控制肥大细胞与其他炎症细胞相互关系的生物活性分子作用机制的基本问题进行了研究，主要集中在以下三点。1. 肥大细胞组胺合成的研究。(1)从小鼠肥大细胞瘤P-815细胞中分离l -组氨酸脱羧酶，测定其cdna来源的氨基酸序列。(2)两种兴奋剂对培养肥大细胞中l -组氨酸脱羧酶及其mRNA的合成有协同作用；地塞米松和12- o -十四烷基酚13-醋酸酯，或二丁基cAMP和钙离子载体A23187。(3)可能存在l -组氨酸脱羧酶的翻译后加工。2. 肥大细胞与内皮细胞或血小板相互作用的研究。(1)肥大细胞代谢产物DELTA^<12>-PGJ_2抑制内皮细胞的生长，但诱导内皮细胞血红素加氧酶31kDa蛋白的表达。(2)内皮细胞PGI_2通过gs偶联腺苷酸环化酶刺激cAMP合成。(3)凝血酶或atp诱导的肥大细胞瘤细胞内PGI_2受体动员Ca_<2+>的差异调控。(4)利用不可逆特异性光亲和探针对肥大细胞瘤细胞和血小板中的PGI_2受体进行了鉴定。3. 前列腺素E受体亚型结构与功能的研究。以及它们在不同组织中的mRNA表达。(1)小鼠前列腺素E受体EP_3亚型和EP_2亚型cdna的克隆与表达。(2)通过选择性剪接获得了两个不同c端结构域的PGE受体EP_3亚型亚型的功能cDNA。两种形式决定了受体激动剂诱导脱敏的配体结合亲和力和敏感性。