CORE--ANALYTICAL BIOCHEMISTRY LABORATORY

肾微粒体细胞色素P-450在体内外的催化作用 花生四烯酸的氧化代谢产物包括 环氧二十碳三烯酸、19-和20-羟基二十碳四烯酸 酸(19-和20-羟基-AA)。这一分支的潜在意义 花生四烯酸对肾功能的级联作用由a)提出 它的一些代谢物具有很强的生物活性，b) 内源性的，对映体选择性的，形成内源性内源EET的演示，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 4A2基因的优势表达 高血压(SHR)大鼠。最近，EETS演示为 人体肾脏和尿液的内源性成分及其增加 妊娠高血压综合征患者的尿液排泄量表明 途径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 以实验的方式探讨这些重要问题，并支持 项目1-6，核心B将采用标准和既定的方法 二十烷类化合物的提取、纯化、高效液相、GC/MS、蛋白质免疫印迹 并且，对DNA进行纯化和整理，以进行鉴定， 酶及其代谢物的表征和定量 花生四叠石级联的这一分支。

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.