CORE--ANALYTICAL BIOCHEMISTRY LABORATORY

肾微粒体细胞色素 P-450 催化体外和体内 花生四烯酸的氧化代谢产物包括 环氧二十碳三烯酸 (EET)、19- 和 20- 羟基二十碳四烯酸 酸（19-和20-OH-AA）。 该分支的潜在意义 a) 表明花生四烯酸级联对肾功能的影响 其一些代谢物的有效生物活性，b) 演示内源性、对映选择性、EET 的形成，c) 通过饮食调节所涉及的酶和/或其代谢物 盐负荷、DOCA 给药或实验性高血压， d) 细胞色素 P-450 4A2 基因在 高血压（SHR）大鼠。 最近，EET 的示范 人肾和尿液的内源性成分及其增加 妊娠期高血压引起的尿排泄表明， 该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 通过实验解决这些重要问题，并支持 项目 1-6，核心 B 将应用以下标准和既定方法 类二十烷酸提取、纯化、HPLC、GC/MS、蛋白质免疫印迹 以及 cDNA 纯化和鉴定文件， 酶和代谢物的表征和定量 花生四烯酸级联的这个分支。

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.