Role of Icosaniods in Renal Function

二十烷酸在肾功能中的作用

• 批准号: 6810248
• 负责人: JORGE CAPDEVILA
• 金额: $ 169.44万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6810248
• 关键词: clinical research; cytochrome P450; eicosanoid metabolism; kidney function; renal tubular transport

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.

肾脏在控制体液体积和组成中起关键作用，并且肾小管和/或血流动力学功能障碍是诸如中风、高血压和糖尿病等疾病的常见特征。P450（P450）花生四烯酸（AA）单加氧酶是AA级联的成员，生物合成几种区域异构的羟基和环氧-AA衍生物，其能够调节肾血流动力学、肾小管转运和肾血管反应性。 体外P450基因亚型特异性表型的细胞和动物模型，包括P450敲除小鼠品系，已经证实了这些酶的功能重要性，促进了对其生理作用的研究，并提供了对其代谢产物作用机制的见解。通过建立在这些和以前的研究基础上，本申请提出：a）整合的分子、生物化学和功能方法，以进一步表征生理学上的生物学特性， 肾AA单加氧酶的重要性，和B）开始分析其在人类疾病的病理生理学中的作用。为了这些目的，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，重组DNA和分子遗传学方法在细胞，器官和整个动物水平的P450异构体特异性表型的研究。功能研究将包括：a）生物化学 记录细胞、器官或整个动物AA代谢中的P450亚型特异性变化，B）表征细胞表型，包括激素反应、信号传导机制和离子通量和通道效应，c）鉴定和表征血液动力学和/或肾小管效应，和d）携带破坏的P450基因的小鼠品系用于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的用途 和/或病理生理学。将鉴定、克隆、表达和表征功能相关的鼠和/或大鼠P450的人类同源物。高通量测序将用于确定P450基因结构/表达的改变与高血压等疾病的病理生理学之间的潜在关联。我们的长期目标是从分子水平上了解P450类花生酸在肾脏生理中的作用， 和作用部位以及与人类疾病的相关性。这些都是制定有意义的办法所必需的： a）病理生理学意义的明确定义，和B）未来的药理学靶向，以及临床诊断和干预。