Renal, Non-cyclooxygenase Arachidonate Metabolism

细胞色素P450(P450)花生四烯酸(AA)单加氧酶的研究揭示了该酶系统在细胞和器官生理学中的新的重要功能。P450衍生的代谢物中最广泛的特征是环氧酸和omega/omega-1羟基花生四烯酸，它们与肾小管钠和水运输、肾脏血流动力学和肾血管反应性的调节有关。此外，P450功能和/或表达的基因控制变化与全身血压控制之间的关联表明，这些酶在肾脏和身体的动态平衡以及高血压的病理生理学中发挥着重要作用。然而，肾脏P450的生理意义及其代谢产物的作用部位和作用方式仍有待明确，需要建立先进的P450 AA单加氧酶同工酶功能表型的实验模型。项目2结合本计划项目的细胞和器官生理学部分，建议利用分子方法开发和生物分子表征P450异构体依赖的功能和/或功能障碍的模型。P450基因的破坏和/或过度表达将用于整合的功能 特异性肾AA的意义及功能作用的生化分析(S) 环氧合酶和omega/omega-1羟基酶。我们将结合使用分析、 生化和重组DNA技术分析P450基因依赖的二十烷类化合物生物合成、AA代谢和P450异构体器官表达和调控的变化。该项目的长期目标是提供对肾脏P450二十烷类化合物生物学意义和作用方式的分子理解。这些重要问题的答案对于制定有意义的方法来明确定义：a)它们的生理意义，b)与高血压等人类疾病的相关性，以及为未来的药理学和/或临床干预制定合理的策略，是必要的。

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.