Role of Icosaniods in Renal Function

肾脏在控制体液容量和成分方面起着关键作用，而肾小管和/或血液动力学功能障碍是中风、高血压和糖尿病等疾病的常见特征。CYP P450(P450)花生四烯酸(AA)单加氧酶是AA级联中的一员，它生物合成几种区域异构体羟基和环氧AA衍生物，能够调节肾脏血流动力学、肾小管转运和肾血管反应性。 体外培养。P450基因异构体特异性表型的细胞和动物模型，包括P450基因敲除小鼠品系，已经证实了这些酶的功能重要性，促进了对它们的生理作用的研究，并对它们的代谢产物的作用机制提供了深入的了解。通过建立在这些和以前的研究的基础上，本申请提出：a)一种综合的分子、生化和功能方法来进一步表征生理 肾脏AA单加氧酶的意义，以及b)启动其在人类疾病的病理生理学中的作用的分析。为此，我们将继续利用合成代谢物、异构体特异性抑制剂、克隆的cDNA和/或基因，以及重组DNA和分子遗传学方法，在细胞、器官和整个动物水平上研究P450异构体的特异性表型。功能研究将包括：a)生化 记录P450亚型在细胞、器官或整个动物AA代谢中的特异性变化b)细胞表型的表征，包括激素反应、信号机制以及离子流量和通道效应，c)血流动力学和/或管状效应的鉴定和表征，以及d)利用携带被破坏的P450基因的小鼠品系将基因特异性细胞和/或器官表型整合到整个动物生理学中 和/或病理生理学。功能相关的小鼠和/或大鼠P450的人类同源物将被鉴定、克隆、表达和鉴定。高通量测序将用于确定P450基因结构/表达变化与高血压等疾病的病理生理学之间的潜在联系。我们的长期目标是提供对P450二十烷酸类化合物在肾脏生理中的作用(S)的分子理解，及其机制 和作用地点，以及与人类疾病的相关性。这些都是制定有意义的方法以实现以下目标所必需的： A)病理生理意义的明确定义，以及b)未来的药物靶向，以及临床诊断和干预。

The kidney plays a key role in the control of body fluid volume and composition, and tubular and/or hemodymic dysfunction are common features of diseases such as stroke, hypertension, and diabetes. The CYP P450 (P450) arachidonic acid (AA) monooxygenase, a member of the AA cascade, biosynthesizes several regioisomeric hydroxyand epoxy-AA derivatives capable of modulating renal hemodynamics, tubular transport, and renovascular reactivity in vitro. Cell and animal models of P450 gene-isoform specific phenotypes, including P450 knockout mice strains, have confirmed the functional importance of these enzymes, facilitated studies of their physiological roles, and provided insights into the mechanism of action of their metabolites. By building upon these and previous studies, this application proposes: a) an integrated molecular, biochemical, and functional approach to further characterize the physiological significance of the renal AA monooxygenase, and b) to initiate the analysis of its role in the pathophysiology of human disease. For these purposes, we will continue to utilize synthetic metabolites, isoform specific inhibitors, cloned cDNAs and/or genes, and recombinant DNA and molecular genetics approaches for the study of P450isoform specific phenotypes at the cellular, organ and whole animal levels. Functional studies will include: a) biochemical documentation of P450 isoform specific changes in cell, organ, or whole animal AA metabolism b) characterization of cellular phenotypes including hormonal responses, signaling mechanisms, and ion flux and channel effects, c) the identification and characterization of hemodynamic and/or tubular effects, and d) the use of mice strains carrying disrupted P450 genes for the integration of gene specific cell and/or organ phenotypes into whole animal physiology and/or pathophysiology. The human homologues of functionally relevant murine and/or rat P450s will be identified, cloned, expressed, and characterized. High throughput sequencing will be used to define potential associations between alterations in P450 gene structure/expression and the pathophysiology of diseases such as hypertension. Our long term goals are to provide a molecular understanding of role(s) of P450 eicosanoids in renal physi ological, their mechanism and site of action, and relevance to human disease. These are needed for the development of meaningful approaches for: a) the unequivocal definition of pathophysiological significance, and b) future pharmacological targeting, and clinical diagnosis and intervention.