Cytochrome P450 Derived Eicosanoids and Inflammation

摘要： 炎症是一个基本的过程，在许多疾病的发病机制中起着不可或缺的作用 人类的国家。核因子κ B（NF-B）是炎症反应的中心介质， 细胞因子、趋化因子和细胞粘附分子表达的转录激活。因此，委员会认为， 鉴定旨在抑制这种病理过程的新策略提供了实质性的治疗方法， 潜力花生四烯酸被来自于花生四烯酸的细胞色素P450（P450）环氧合酶氧化代谢。 在肝和肝外组织中，CYP 2 J和CYP 2C亚家族与环氧二十碳三烯酸（E3）结合。的 可溶性环氧化物水解酶（sEH）可将雌二醇快速水解为活性较低的二羟基二十碳三烯酸 （DHE）。花生四烯酸也被代谢为20-羟基二十碳四烯酸（20-HETE）， 来自CYP 4A和CYP 4F亚家族的羟化酶。最近的证据表明，CYP衍生的 雌二醇和20-HETE分别具有抗炎和促炎作用。然而， CYP介导的类花生酸代谢对体内炎症的调节作用尚未得到严格的研究 表征了我们假设，β-环氧合酶-和β-环氧合酶-之间的功能平衡， 羟化酶介导的花生四烯酸代谢是调节NF-κ B介导的 体内炎症反应，并调节这种平衡，有利于β-环氧合酶途径 提供了巨大的治疗潜力。本提案的主要目标是：（1）确定 肝脏和肝外CYP介导的类花生酸代谢的炎症反应，（2）定义 EET的生物合成和sEH介导的EET水解在EET合成中的功能作用 调节炎症，和（3）表征表氧化酶和β-环氧化酶的相对影响， 羟化酶途径调节体内炎症反应。该项目将利用新型转基因 以及基因敲除小鼠和药理学工具来操纵CYP介导的体内类花生酸代谢， 同时使用已建立的分子生物学来表征NF-κ B介导的炎症反应， 分析技术。总的来说，这一系列新颖的实验将定义 表氧化酶和羟化酶衍生的20-HETE对肝和肝细胞凋亡的调节作用 体内肝外炎症反应，并促进新的抗炎药的开发 具有潜在的治疗应用于人类多种疾病状态的策略。

ABSTRACT: Inflammation is a fundamental process which plays an integral role in the pathogenesis of numerous disease states in humans. Nuclear factor kappa B (NF-¿B) is a central mediator of the inflammatory response via transcriptional activation of cytokine, chemokine and cellular adhesion molecule expression. Consequently, identification of novel strategies aimed at inhibition of this pathological process offers substantial therapeutic potential. Arachidonic acid is oxidatively metabolized by cytochrome P450 (CYP) epoxygenases from the CYP2J and CYP2C subfamilies to epoxyeicosatrienoic acids (EETs) in hepatic and extra-hepatic tissue. The EETs are rapidly hydrolyzed by soluble epoxide hydrolase (sEH) to less active dihydroxyeicosatrienoic acids (DHETs). Arachidonic acid is also metabolized to 20-hydroxyeicosatetraenoic acid (20-HETE) by CYP ¿- hydroxylases from the CYP4A and CYP4F subfamilies. Recent evidence has demonstrated that CYP-derived EETs and 20-HETE possess anti- and pro-inflammatory effects, respectively. However, the contribution of CYP-mediated eicosanoid metabolism to the regulation of inflammation in vivo has not been rigorously characterized. We hypothesize that the functional balance between CYP epoxygenase- and CYP ¿- hydroxylase-mediated arachidonic acid metabolism is integral to the regulation of NF-¿B-mediated inflammatory responses in vivo, and modulation of this balance in favor of the CYP epoxygenase pathway offers substantial therapeutic potential. The primary objectives of this proposal are to: (1) define the impact of the inflammatory response on hepatic and extra-hepatic CYP-mediated eicosanoid metabolism, (2) define the functional role of CYP epoxygenase-mediated EET biosynthesis and sEH-mediated EET hydrolysis in the regulation of inflammation, and (3) characterize the relative impact of CYP epoxygenase and CYP ¿- hydroxylase pathway modulation on inflammatory responses in vivo. This project will utilize novel transgenic and knock-out mice and pharmacological tools to manipulate CYP-mediated eicosanoid metabolism in vivo, while characterizing NF-¿B-mediated inflammatory responses using established molecular biology and analytical techniques. Collectively, this series of novel experiments will define the mechanistic contribution of CYP epoxygenase-derived EETs and CYP ¿-hydroxylase-derived 20-HETE to the regulation of hepatic and extra-hepatic inflammatory responses in vivo, and facilitate the development of new anti-inflammatory strategies with potential therapeutic application to numerous disease states in humans.