Renal, Non-cyclooxygenase Arachidonate Metabolism

肾脏、非环氧合酶花生四烯酸代谢

• 批准号: 6813193
• 负责人: JORGE CAPDEVILA
• 金额: $ 16.23万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6813193
• 关键词: arachidonate; cytochrome P450; dietary sodium; disease /disorder model; eicosanoid metabolism; enzyme activity; gas chromatography mass spectrometry; gene targeting; genetically modified animals; immunoelectrophoresis; kidney function; kidney metabolism; laboratory mouse; microsomes; nucleic acid hybridization; oxidation; oxygenases; phenotype; protein isoforms; protein purification; recombinant proteins; tissue /cell culture; transfection /expression vector

The studies of the cytochrome P450 (P450) arachidonic acid (AA) monooxygenase, now established as a major pathway for the bioactivation of endogenous AA, uncovered new and important functional roles for this enzyme system in cell and organ physiology. The most extensively characterized of the P450-derived metabolites, the epoxy- and the omega/omega-1 hydroxy-arachidonic acid, have been implicated in the regulation of tubular sodium and water transport, renal hemodynamics, and renovascular reactivity. Furthermore, associations between genetically controlled alterations in P450 function and/or expression, and the control of systemic blood pressures suggest important roles for these enzymes in kidney and body homeostasis, and in the pathophysiology of hypertension. However, the physiological significance of renal P450 and the site and mode of action of its metabolites remains to be unequivocally defined, and there is a need for advanced experimental models of P450 AA Monooxygenase, isoform-dependent, functional phenotypes. Project 2, in conjunction with the cell and organ physiology components of this Program Project, proposes to utilize molecular approaches for the development and bio-molecular characterization of models of P450 isoform-dependent function and/or dysfunction. P450 gene disruption and/or overexpression will be employed for the integrated functional and biochemical analysis of the significance and the functional role(s) of specific kidney AA epoxygenase and omega/omega-1 hydroxylases. We will apply a combination of analytical, biochemical, and recombinant DNA techniques to the analysis of P450 gene-dependent changes in eicosanoid biosynthesis, AA metabolism, and P450 isoform organ expression and regulation. The long term goals of this project are to provide a molecular understanding of renal P450 eicosanoid biological significance and mode of action. The answers to these important questions are needed for the development of meaningful approaches to the unequivocal definition of: a) their physiological significance, b) relevance to the human diseases such as hypertension, and for the development of rational strategies for future pharmacological and/or clinical intervention.

细胞色素P450 （P450）花生四烯酸（AA）单加氧酶现已被确定为内源性AA生物活化的主要途径，其研究揭示了该酶系统在细胞和器官生理中的新的重要功能作用。最广泛表征的p450衍生代谢物，环氧-和omega/omega-1羟基花生四烯酸，与管状钠和水运输、肾脏血流动力学和肾血管反应性的调节有关。此外，基因控制的P450功能和/或表达改变与控制全身血压之间的关联表明，这些酶在肾脏和身体稳态以及高血压的病理生理中起着重要作用。然而，肾脏P450的生理意义及其代谢物的作用部位和方式仍然没有明确的定义，并且需要先进的P450 AA单加氧酶的实验模型，同型依赖性，功能性表型。项目2，结合本项目的细胞和器官生理学组成部分，提出利用分子方法开发和生物分子表征P450亚型依赖的功能和/或功能障碍模型。P450基因破坏和/或过表达将用于整合功能