Cytochrome P450 Derived Eicosanoids and Inflammation

细胞色素 P450 衍生的类二十烷酸与炎症

• 批准号: 8304582
• 负责人: CRAIG R LEE
• 金额: $ 2.85万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8304582
• 关键词: Acids; Acute; Alkane 1-monooxygenase; Anabolism; Anti-Inflammatory Agents; Anti-inflammatory; Arachidonic Acids; Attenuated; Biochemical; Biological Assay; Blood Vessels; CYP2J2 gene; Cardiovascular system; Cell Adhesion; Cell Adhesion Molecules; Chemosensitization; Cytochrome P450; Data; Development; Disease; Eicosanoids; Endothelial Cells; Enzyme-Linked Immunosorbent Assay; Epoxide hydrolase; Equilibrium; Exhibits; Functional disorder; Generations; Genetic Transcription; Health; Hepatic; Hepatic Tissue; Histologic; Human; Hydrolysis; Hydroxyeicosatetraenoic Acids; Immunoblotting; In Vitro; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Knockout Mice; Laboratories; Leukocytes; Lipopolysaccharides; Maintenance; Mediating; Mediator of activation protein; Metabolic; Metabolism; Mixed Function Oxygenases; Molecular; Molecular Biology; Mus; NF-kappa B; Nitric Oxide Synthase; Pathogenesis; Pathologic Processes; Pathway interactions; Peroxidases; Play; Process; Prostaglandin-Endoperoxide Synthase; Regulation; Relative (related person); Research Project Grants; Reverse Transcriptase Polymerase Chain Reaction; Role; Series; Signal Transduction; Stimulus; Techniques; Testing; Therapeutic; Tissues; Transcriptional Activation; Transgenic Mice; Transgenic Organisms; Vascular Endothelial Cell; Vasoconstrictor Agents; Wild Type Mouse; Work; Xenobiotic Metabolism; attenuation; base; chemokine; cytochrome P-450 CYP2C subfamily; cytokine; defined contribution; eicosanoid metabolism; in vivo; inhibitor/antagonist; novel; novel strategies; novel therapeutics; overexpression; promoter; protein expression; research study; tool; transcription factor; vascular inflammation

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: Inflammation is a fundamental process which plays an integral role in the pathogenesis of numerous diseases in humans. Completion of this research project will define the contribution of cytochrome P450-mediated eicosanoid metabolism to the regulation of inflammation in vivo, characterize the underlying mechanisms, and facilitate the development of novel therapeutic strategies aimed at inhibition of inflammation.

描述（由申请人提供）：炎症是一个基本过程，在人类多种疾病状态的发病机制中发挥着不可或缺的作用。核因子 kappa B (NF-κB) 是通过转录激活细胞因子、趋化因子和细胞粘附分子表达而产生炎症反应的中心介质。因此，识别旨在抑制这种病理过程的新策略提供了巨大的治疗潜力。花生四烯酸在肝脏和肝外组织中被 CYP2J 和 CYP2C 亚家族的细胞色素 P450 (CYP) 环氧化酶氧化代谢为环氧二十碳三烯酸 (EET)。 EET 被可溶性环氧化物水解酶 (sEH) 快速水解为活性较低的二羟基二十碳三烯酸 (DHET)。花生四烯酸还被 CYP4A 和 CYP4F 亚家族的 CYP α-羟化酶代谢为 20-羟基二十碳四烯酸 (20-HETE)。最近的证据表明，CYP 衍生的 EET 和 20-HETE 分别具有抗炎和促炎作用。然而，CYP 介导的类二十烷酸代谢对体内炎症调节的贡献尚未得到严格表征。我们假设 CYP 环氧化酶和 CYPβ-羟化酶介导的花生四烯酸代谢之间的功能平衡对于体内 NF-κB 介导的炎症反应的调节是不可或缺的，并且有利于 CYP 环氧化酶途径的这种平衡的调节提供了巨大的治疗潜力。该提案的主要目标是：（1）定义炎症反应对肝脏和肝外 CYP 介导的类二十烷酸代谢的影响，（2）定义 CYP 环氧化酶介导的 EET 生物合成和 sEH 介导的 EET 水解在炎症调节中的功能作用，以及（3）表征 CYP 环氧化酶和 CYP 的相对影响 β-羟化酶途径对体内炎症反应的调节。该项目将利用新型转基因和基因敲除小鼠以及药理学工具来操纵 CYP 介导的类二十烷酸体内代谢，同时使用已建立的分子生物学和分析技术来表征 NF-κB 介导的炎症反应。总的来说，这一系列新颖的实验将确定 CYP 环氧化酶衍生的 EET 和 CYP β-羟化酶衍生的 20-HETE 对体内肝脏和肝外炎症反应调节的机制贡献，并促进新的抗炎策略的开发，对人类多种疾病状态具有潜在的治疗应用。公共卫生相关性：炎症是一个基本过程，在人类多种疾病的发病机制中发挥着不可或缺的作用。该研究项目的完成将明确细胞色素 P450 介导的类二十烷酸代谢对体内炎症调节的贡献，表征潜在机制，并促进旨在抑制炎症的新型治疗策略的开发。