NADPH-DEPENDENT METABOLISM OF ARACHIDONIC ACID

细胞色素P450(P450)依赖的花生四烯酸(AA)单加氧酶， 现在被认为是生物医学上重要的AA的第三个分支 卡斯卡德。P450二十烷类化合物中最广泛的特征是环氧基- 和19/20-羟基花生四烯酸，被认为与控制 血管反应性、离子通量、激素信号和血管病变的病理生理学 实验性高血压。然而，由于缺乏合适的分子 这些工具的作用机制和生理意义 代谢物仍有待确定。假设是：a) 细胞内合成特定的P450二十烷类化合物提供了重要的 细胞和器官功能的脂质介体，B)氧化代谢的作用 在细胞/器官P450二十烷类激素水平的控制中发挥重要作用，因此， 生物功能和效力，以及C)配体激活的过氧化物体 增殖物激活受体α(PPARpha)参与 P450二十烷类化合物和脂肪酸氧化代谢的协调调节。 由于二十烷类化合物的信息内容、信号特性和 生物作用由酶、异构体特异性、区域和 AA分子模板的立体选择性氧合，该项目将 利用重组DNA、生化和生物分析的组合 技术：a)区域选择AA的设计和施工 单加氧酶及其在培养细胞中的表达，b)文献 细胞P450二十烷类化合物的生物合成和代谢，c)研究 依赖于c DNA的细胞表型，以及d)功能和生化 过氧化体-β-氧化与微粒体的作用分析(S) Omega/Omega-1羟化在P450-二十烷类化合物代谢和功能中的作用。这个 总体目标是提供生物化学和分子方面的描述 该通路在器官/身体生理学和/或病理生理学中的作用(S)。 已有文献记载的P450二十烷类化合物的细胞和系统效应及其 潜在的生物医学意义继续为 这种代谢途径的研究。这些问题的实验答案 这些假设所提出的假设需要：a)明确定义 P450AA的生理和/或病理生理学意义 单加氧酶，以及b)为未来制定有效的战略 药理学和/或临床干预。

The cytochrome P450 (P450)-dependent arachidonic acid (AA) monooxygenases, are now recognized as the third branch of the bio-medically important AA cascade. The most extensively characterized of the P450 eicosanoids, the epoxy- and the 19/20-hydroxy arachidonic acids, have been implicated in the control of vascular reactivity, ion fluxes, hormonal signaling and the pathophysiology of experimental hypertension. However, due to paucity of appropriate molecular tools, the mechanisms of action and physiological significance of these metabolites remains to be established. The hypotheses are that: A) the intracellular biosynthesis of specific P450 eicosanoids provides important lipid mediators of cellular and organ function, B) oxidative metabolism plays an important role in the control of cell/organ P450 eicosanoid levels and thus, biological function and potency, and C) the ligand activated peroxisomal proliferator activated receptor alpha (PPARalpha) participates in the coordinated regulation of P450 eicosanoid and fatty acid oxidative metabolism. Inasmuch as eicosanoid informational content, signaling properties, and biological role are controlled by enzymatic, isoform specific, regio-, and stereoselective oxygenations of the AA molecular template, this project will utilize combinations of recombinant DNA, biochemical, and bioanalytical techniques for: a) the design and construction of regioselective AA monooxygenases and their expression in cultured cells, b) the documentation of cellular P450 eicosanoid biosynthesis and metabolism, c) the study of cDNA-dependent cellular phenotypes, and d) the functional and biochemical analysis of the role(s) of peroxisomal beta-oxidation and microsomal omega/omega-1 hydroxylation in P450-eicosanoid metabolism and function. The overall goals are to provide a biochemical and molecular description of the role(s) of this pathway in organ/body physiology and/or pathophysiology. The documented cellular and systemic effects of the P450 eicosanoids and their potential biomedical significance continue to provide the rationale for the studies of this metabolic pathway. The experimental answers to the questions posed by these hypotheses are needed for: a) the unequivocal definition of the physiological and/or pathophysiological significance of the P450 AA monooxygenases, and b) the formulation of effective strategies for future pharmacological and/or clinical intervention.