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 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学中新的重要功能作用。 P450 衍生代谢物中最广泛表征的环氧-和 omega/omega-1 羟基花生四烯酸与肾小管钠和水转运、肾血流动力学和肾血管反应性的调节有关。此外，P450 功能和/或表达的基因控制改变与全身血压控制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学中发挥重要作用。然而，肾脏 P450 的生理意义及其代谢物的作用位点和模式仍有待明确定义，并且需要 P450 AA 单加氧酶、异构体依赖性功能表型的先进实验模型。项目 2 与该计划项目的细胞和器官生理学组成部分相结合，建议利用分子方法来开发 P450 异构体依赖性功能和/或功能障碍模型并对其进行生物分子表征。 P450 基因破坏和/或过度表达将用于集成功能 特定肾脏 AA 的意义和功能作用的生化分析 环氧化酶和 omega/omega-1 羟化酶。我们将结合分析、 生物化学和重组 DNA 技术来分析类二十烷酸生物合成、AA 代谢以及 P450 亚型器官表达和调节中 P450 基因依赖性变化。该项目的长期目标是提供对肾 P450 类二十烷酸生物学意义和作用模式的分子理解。这些重要问题的答案对于开发有意义的方法来明确定义：a）其生理意义，b）与高血压等人类疾病的相关性，以及为未来的药理学和/或临床干预。