CYTOCHROME P450 DEPENDENT ARACHIDONIC ACID METABOLISM

描述（来自申请人摘要的逐字描述）： 细胞色素P450（CYP）催化花生四烯酸（AA）代谢是 ω-羟基化以形成20-羟基二十碳四烯酸（20-HETE），和 产生区域和立体异构的环氧二十碳三烯酸的环氧化反应 （埃塞尔比）。雌二醇进一步被环氧化物水解酶代谢为 相应的二羟基二十碳三烯酸（DHEA）。这些CYP衍生物 类二十烷酸是令人感兴趣的，因为它们是许多植物的内源性成分。 组织和神经系统产生广泛的生物学效应。小区特定 酶的表达模式、相对丰度和活性 催化这些反应将是细胞内 这些类二十烷酸的作用。在肾脏中，CYP衍生的类花生酸具有强效的 对肾血管张力和肾小管离子转运的影响， 与血压调节有关。的总体目标 提出的研究是了解控制 CYP催化的AA在大鼠肾脏中的代谢及肾脏保护的重要性 类花生酸在血压调节中的作用的具体目标 建议的研究是（1）检查的机制基础， 抑制AA ω-羟化酶活性的抗高血压作用。 具体而言，我们将确定的异构体特异性和效力 基于机制的抑制剂的β-AA ω-羟化酶，并评估其 对血压和血管张力的影响;（2）确定 CYP 4 E亚型对肾脏AA代谢的影响。CYP 4F表达将定位于 将表征大鼠肾脏和CYP 4F亚型的AA代谢; (3)分离大鼠肾脏中表达的主要CYP 2 J环氧合酶。的 编码CYP 2 J2免疫反应性蛋白的cDNA在小鼠中过表达， 通过表达克隆鉴定自发性高血压大鼠肾脏 和功能特征;（4）检查EET的调节 在大鼠肾脏中水解。具体来说，我们将确定生物化学 EET在大鼠肾微粒体中水解的基础和可溶性 环氧化物水解酶对血压和肾类花生酸形成的抑制。 这些研究的结果将有助于全面了解 表达和调节主要的环氧化物和环氧化物水解酶 参与AA代谢。这些研究的一个长期目标是开发 用于调节肾类花生酸的新靶向治疗剂 生产和血压。应用这些原则进行管理 预期在其他组织中会形成类二十烷酸。

DESCRIPTION (Verbatim from Applicant's Abstract): The two major pathways of cytochrome P450 (CYP)-catalyzed arachidonic acid (AA) metabolism are omega-hydroxylation to form 20-hydroxyeicosatetraenoic acid (20-HETE) and epoxidation which produces regio- and stereoisomeric epoxyeicosatrienoic acids (EETs). The EETs are further metabolized by epoxide hydrolases to the corresponding dihydroxyeicosatrienoic acids (DHETs). These CYP-derived eicosanoids are of interest since they are endogenous constituents of numerous tissues and posses a wide array of biological effects. The cell specific pattern of expression, relative abundance, and activity of the enzymes catalyzing these reactions will be a major determinant of the intracellular effect of these eicosanoids. In the kidney, CYP-derived eicosanoids have potent effects on renal vascular tone and tubular ion transport and have been implicated in the regulation of blood pressure. The overall goal of the proposed studies is to understand the molecular mechanisms controlling CYP-catalyzed AA metabolism in the rat kidney and the importance of renal CYP eicosanoid levels in blood pressure regulation. The specific aims of the proposed studies are (1) to examine the mechanistic basis of the antihypertensive effect of inhibition of AA omega-hydroxylase activity. Specifically, we will determine the isoform-specificity and potency of mechanism-based inhibitors of CYP AA omega-hydroxylases and evaluate their effect on blood pressure and vascular tone; (2) to determine the contribution of CYP4E isoforms to renal AA metabolism. CYP4F expression will be localized in the rat kidney and AA metabolism by the CYP4F isoforms will be characterized; (3) to isolate the major CYP2J epoxygenase expressed in the rat kidney. The cDNA encoding the CYP2J2 immunoreactive protein overexpressed in the spontaneously hypertensive rat kidney will be identified by expression cloning and functionally characterized; and (4) to examine the regulation of EET hydrolysis in the rat kidney. Specifically, we will determine the biochemical basis of EET hydrolysis in rat renal microsomes and the effect of soluble epoxide hydrolase inhibition on blood pressure and renal eicosanoid formation. The findings from these studies will lead to a comprehensive understanding of the expression and regulation of the major CYP and epoxide hydrolase enzymes involved in AA metabolism. A long term goal of these studies is to develop novel targeted therapeutics for the regulation of renal CYP eicosanoid production and blood pressure. Application of these principles for regulating CYP eicosanoid formation in other tissues is anticipated.