CORE--ANALYTICAL BIOCHEMISTRY LABORATORY

核心--分析生化实验室

• 批准号: 6270658
• 负责人: CAPDEVILA JORGE H
• 金额: $ 5.6万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-01 至 1998-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6270658
• 关键词: arachidonate; biochemistry; biomedical facility; eicosanoid metabolism; gas chromatography mass spectrometry; genetic library; high performance liquid chromatography; radioimmunoassay

Renal microsomal cytochrome P-450 catalyzes the in vitro and in vivo oxygenated metabolism of arachidonic acid to products that include epoxyeicosatrienoic acids (EETs), 19- and 20-hydroxyeicosatetraenoic acids (19- and 20-OH-AA). The potential significance of this branch of the arachidonate cascade to renal function has been suggested by a) the potent biological activities of some of its metabolites, b) the demonstration of endogenous, enantioselective, formation of EETs, c) the regulation of the enzymes involved and/or of its metabolites by dietary salt loading, DOCA administration or by experimental hypertension and, d) the preferential expression of the cytochrome P-450 4A2 gene in hypertensive (SHR) rats. More recently, the demonstration of EETs as endogenous constituents of human kidney and urine and their increased urinary excretion during pregnancy induced hypertension suggest that this pathway may fulfill a yet unidentified role in human kidney physiology and/or pathophysiology. As part of a multidisciplinary effort to approach experimentally these important questions and, in support of projects 1-6, Core B will apply standard and established methods of eicosanoid extraction, purification, HPLC, GC/MS, protein immunoblotting and, cDNA purification and documentation to the identification, characterization and quantification of the enzymes and the metabolites of this branch of the arachidonate cascade.

肾微粒体细胞色素P-450在体外和体内催化 花生四烯酸的氧化代谢产物，包括 环氧二十碳三烯酸（ESTs），19-和20-羟基二十碳四烯酸 酸（19-和20-OH-AA）。 这个分支的潜在意义 花生四烯酸对肾功能的级联反应已经由以下提出：a） 它的一些代谢产物的有效生物活性，B） 证明内源性的、对映选择性的、E2的形成，c） 饮食对相关酶和/或其代谢物的调节 盐负荷、DOCA给药或实验性高血压， d）细胞色素P-450 4A 2基因在大肠杆菌中的优先表达， 高血压（SHR）大鼠。 最近，埃克塞特的演示 人体肾脏和尿液的内源性成分及其增加 妊娠高血压综合征期间的尿排泄表明， 这一通路可能在人类肾脏生理学中发挥尚未确定的作用 和/或病理生理学。 作为多学科努力的一部分， 实验性地探讨这些重要问题，并支持 项目1-6，核心B将采用标准和既定的方法， 类花生酸提取，纯化，HPLC，GC/MS，蛋白质免疫印迹 以及，cDNA纯化和文献鉴定， 酶和代谢物的表征和定量 花生四烯酸级联反应的分支。