ETHANOL TOLERANCE AND DEPENDENCE IN VITRO

体外乙醇耐受性和依赖性

• 批准号: 3108910
• 负责人: ELLIOTT RICHELSON
• 金额: $ 12.97万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1980
• 资助国家: 美国
• 起止时间: 1980-04-01 至 1989-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3108910
• 关键词: adenylate cyclase; angiotensin II; arachidonate; bradykinin; clone cells; cyclic AMP; cyclic GMP; drug abuse; drug tolerance; ethanol; fibroblasts; guanylate cyclase; high performance liquid chromatography; histamine; inositol phosphates; membrane activity; neuroblastoma; neuropharmacology; phosphatidylinositols; prostaglandin E; radioimmunoassay; synapses; thrombin

The locus of ethanol's pharmacological and addictive effects may be at the level of the interaction between a neurotransmitter and its receptor in the synapse. Therefore this proposal focuses on the effects of acute and chronic ethanol on the functioning of certain receptors which have cyclic AMP and cyclic GMP as putative second messengers. Cultured human skin fibroblasts with prostaglandin and Beta-adrenergic receptors mediating cyclic AMP synthesis and murine neuroblastoma cells (clone N1E-115) with several receptors (muscarinic, histamine H1, angiotensin 11, bradykinin, neurotensin and thrombin) mediating cyclic GMP synthesis are being used as model systems for this work. Studies with human skin fibroblasts will determine whether results found in animal studies (including our work with murine neuroblastoma cells) are applicable to human receptors. Studies with murine neuroblastoma cells may help elucidate the mechanism whereby ethanol depletes brain levels of cyclic GMP. The prostaglandin and Beta-adrenergic receptors of human skin fibroblasts will be thoroughly characterized by biological assay (cyclic AMP production) and by radioligand binding assay prior to the studies with ethanol. The cyclic GMP studies with neuroblastoma cells will follow-up on our data showing a mixed type of inhibition (competitive and noncompetitive) by ethanol of receptor-mediated cyclic GMP synthesis and an adaptation to this effect of ethanol upon chronic exposure of cells to this alcohol. Since receptor-mediated cyclic GMP synthesis involves the turnover of phospholipids, effects of ethanol on the release of inositol phosphates and arachidonate acid will be studied.

乙醇的药理作用和成瘾作用可能位于 神经递质与其受体之间相互作用的水平 突触。 因此，该提案重点关注急性和 长期乙醇对某些具有周期性受体功能的影响 AMP 和环 GMP 作为假定的第二信使。 培养的人类皮肤 前列腺素和β-肾上腺素能受体介导的成纤维细胞 环AMP合成和鼠神经母细胞瘤细胞（克隆N1E-115） 几种受体（毒蕈碱、组胺 H1、血管紧张素 11、缓激肽、 神经降压素和凝血酶）介导环 GMP 合成被用作 这项工作的模型系统。 对人类皮肤成纤维细胞的研究将 确定动物研究（包括我们与 鼠神经母细胞瘤细胞）适用于人类受体。 研究 小鼠神经母细胞瘤细胞可能有助于阐明其中的机制 乙醇会消耗大脑中环鸟苷酸的水平。 前列腺素和 人体皮肤成纤维细胞的β-肾上腺素受体将被彻底 通过生物测定（环 AMP 产生）和 在用乙醇进行研究之前进行放射性配体结合测定。 循环的 对神经母细胞瘤细胞的 GMP 研究将根据我们的数据进行后续研究，显示 乙醇的混合型抑制（竞争性和非竞争性） 受体介导的环 GMP 合成以及对这种效应的适应 细胞长期暴露于这种酒精中时产生乙醇。 自从 受体介导的环 GMP 合成涉及 磷脂、乙醇对磷酸肌醇释放的影响和 将研究花生四烯酸。