Hormonal Regulation of Blood Pressure

描述(由申请人提供)： 该计划项目资助提案旨在研究细胞色素P450(CYP)衍生的二十烷酸类化合物在血管和肾脏控制血压机制中的功能和调节。该提案侧重于涉及20-羟基二十碳四烯酸(20-HETE)、19(R)和18(R)-HETE衍生的细胞色素P4-2E2E1，以及顺式和反式EET构型的环氧二十碳三烯酸(EETs)的血管和/或肾脏机制。实验策略结合了分子、细胞、分离器官和整体动物方法。拟议的研究活动分为四个项目，由三个核心提供支助。项目1研究了由CYP2E1衍生的二十烷类化合物在调节血管反应性和血压中的作用。血压和血管对收缩刺激的反应性将在接受干预(增加(CYP2E1基因转移)或减少(CYP2E1抑制；CYP2E1缺失小鼠)由CYP2E1派生的二十烷类化合物的合成的干预措施的动物中与血管合成的CYP2E1派生的二十烷类化合物和20-HETE有关。项目2研究了20-HETE作为体内细胞色素P4A2基因转移致高血压大鼠内皮细胞功能障碍和激活的决定因素的作用。将研究血管20-HETE产生、内皮功能障碍指数和血压之间的关系。我们将评估血管内皮细胞与血管内皮细胞20-HETE的作用。20-HEETE诱导内皮功能障碍的分子机制将被阐述。项目3将研究EETs在调节大脑皮层集合管(CCD)钠转运中的作用。依赖EETs的花生四烯酸和腺苷对ENaC的影响将作为钠摄入和环氧合酶表达的函数进行研究。EETs抑制ENaC作用的信号机制将被定义。项目4将研究顺式和反式EETs的合成和释放与压力和氧化应激的关系，检查它们的血管调节作用，并确定顺式和反式EETs在花生四烯酸夸大的肾血管扩张效应中的相对贡献。核心A提供行政支持。核心B负责通过质谱学测量二十烷类化合物。Core C为正、反义CYP基因的转导提供了病毒载体。

DESCRIPTION (provided by applicant): This Program Project Grant proposal aims at investigating the function and regulation of cytochrome P450 (CYP)-derived eicosanoids in relation to vascular and renal mechanism of blood pressure control. The proposal focuses on vascular and/or renal mechanisms involving 20-hydroxyeicosatetraenoic acid (20-HETE), CYP2E1-derived 19(R)- and 18(R)-HETE, and epoxyeicosatrienoic acids (EETs) of both the cis-EET and trans-EET configuration. The experimental strategies combine molecular, cellular, isolated organ and whole animal approaches. The proposed research activities are organized into four projects supported by three cores. Project 1 investigates the role of CYP2E1-derived eicosanoids in the regulation of vascular reactivity and blood pressure. Blood pressure and vascular reactivity to constrictor stimuli will be studied in relation to vascular synthesis of CYP2E1-derived eicosanoids and 20-HETE in animals subjected to interventions that increase (CYP2E1 gene transfer) or decrease (CYP2E1 inhibition; CYP2E1-null mice) the synthesis of CYP2E1-derived eicosanoids. Project 2 investigates the role of 20-HETE as a determinant of endothelial cell dysfunction and activation in rats made hypertensive by in vivo CYP4A2 gene transfer. Relationships will be examined between vascular 20-HETE production, indices of endothelial dysfunction, and blood pressure. The contribution of endothelial versus smooth muscle 20-HETE will be assessed. The molecular mechanisms of 20-HEETE-induced endothelial dysfunction will be addressed. Project 3 will investigate the role of EETs in regulation of Na transport in the cortical collecting duct (CCD). The EETs-dependent effects of arachidonic acid and adenosine on ENaC will be studied as a function of Na intake and epoxygenase expression. The signaling mechanisms underlying the inhibitory action of EETs on ENaC will be defined. Project 4 will study the renal synthesis and release of cis- and trans-EETs in relation to pressure and oxidative stress, examine their vasoregulatory actions, and determine the relative contribution of cis and trans-EETs to the exaggerated renal vasodilatory effect of arachidonic acid in SHR. Core A provides administrative support. Core B is responsible for measurement of eicosanoids by mass spectrometry. Core C provides viral vectors to transfer CYP genes in the sense and antisense orientation.