Role of Cyp2j-epoxygenases, sEH and PPARs in adenosine-induced vascular response

Cyp2j-环氧合酶、sEH 和 PPAR 在腺苷诱导的血管反应中的作用

• 批准号: 8501948
• 负责人: Mohammed A Nayeem
• 金额: $ 35.71万
• 依托单位: WEST VIRGINIA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-01 至 2017-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8501948
• 关键词: Acetylcholine; Adenosine; Adenosine A1 Receptor; Adenosine A2A Receptor; Adenylate Cyclase; Affect; Agonist; Anti-Inflammatory Agents; Anti-inflammatory; Aorta; Bathing; Blood Pressure; Blood Vessels; Cardiovascular Diseases; Cell Line; Cell physiology; Clinical; Coronary artery; Country; Coupling; Cyclic AMP-Dependent Protein Kinases; Data; Development; Down-Regulation; Endothelial Cells; Endothelium; Epoxide hydrolase; Exercise; Exhibits; Experimental Designs; Functional disorder; GTP-Binding Proteins; Genes; Genetic Polymorphism; Genetic Variation; Goals; Human; Hypertension; Image; Individual; Kidney; Link; Measures; Mediating; Mitogen-Activated Protein Kinase Inhibitor; Mus; Myocardial perfusion; Organ; Outcome; PKA inhibitor; PPAR Pathway; Pathway interactions; Patients; Peroxisome Proliferator-Activated Receptors; Pharmaceutical Preparations; Pharmacology; Population; Prevention; Proteins; Publishing; Purinergic P1 Receptors; Receptor Gene; Relaxation; Reporting; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Signal Transduction; Smooth Muscle Myocytes; Technology; Testing; Therapeutic Uses; Transgenic Mice; Vascular Endothelium; Vasodilation; Western Blotting; Wild Type Mouse; base; blood pressure regulation; drinking water; inhibitor/antagonist; innovation; knockout gene; mRNA Expression; novel; overexpression; public health relevance; renal artery; response; tool; vasoconstriction

DESCRIPTION (provided by applicant): Endothelial dysfunction is associated with corresponding changes in vascular tone and increases in contractility, a condition that may cause hypertension in susceptible individuals which may have allelic variants in cyp2j-epoxygenases, soluble epoxide hydrolase (sEH), A2A, and A1 adenosine receptors (A2A AR & A1 AR) genes. These allelic variants may have similarities to our transgenic mice which may regulate vascular tone and blood pressure (BP). Our preliminary data have suggested a possible link between adenosine-induced relaxation and opening of KATP channels through A2A AR-cyp2j-PKA-PPAR? pathway. Also, it is possible that a link may exist between adenosine-induced contraction and closing of KATP channels through A1 AR-sEH-cyp4a- PPAR¿ pathway. A combination of pharmacological tools and transgenic mice would allow us to identify the possible targets as a long term goal to treat population which may have allelic variants leading to hypertension. Therefore, there is a critical need to explore the possible mechanism involving cyp2j2-epoxygenases, sEH/cyp4a, A1 AR/A2A AR, PPAR¿/?, PKA/PKC¿/¿ and KATP channels in adenosine-induced vascular responses. Our central hypothesis is that adenosine induces vascular relaxation and decreases in BP through cyp2j-epoxygenases via A2A AR-cyp2j-PKA-PPAR? signaling leading to opening of KATP channels. On the other hand, adenosine induces vascular contraction and increases in BP through sEH via A1 AR-sEH-cyp4a-PPAR¿ pathway leading to closing of KATP channels. To test this hypothesis, we propose to explore in depth mechanism(s) using A2A AR-/-, A1 AR-/-, cyp2j5-/-, sEH-/-, Tie2-cyp2j2Tr (endothelial-cyp2j2 overexpressed), Tie2-sEHTr (endothelial-sEH overexpressed), wild-type mice, immortal renal endothelial cell line from H-2Kb- tsA58 mouse, mouse aortic endothelial cells (MAEC) and mouse aortic smooth muscle cells (MASMC). Further, we will also explore the possible treatment with sEH inhibitors (AUDA/t-AUCB) in drinking water (or gavage) for A2A AR-/-, cyp2j5-/- and Tie2-sEHTr mice which may have high BP. We will measure BP, and we will use aortas/renal arteries (organ bath/DMT-wire myograph) with treatments (adenosine-receptors agonists & antagonists), cyp-epoxygenases, sEH, adenylyl cyclase, PKC¿/¿/, MAPK and PKA inhibitors, PPAR¿/?, EETs and KATP channel (activators & inhibitors). EETs & DHETs will be analyzed (UPLC-MS/MS). Western blot & RT-PCR will be used for proteins & mRNA expression. We propose 3 specific aims to determine: (1) whether the cyp2j-epxygenases or sEH affects BP, adenosine-induced vascular response and EETs/DHETs; (2) whether the presence or absence of A2A AR affects adenosine-induced vascular response through PPARs via cyp2j-epoxygenases, sEH (3) whether the presence /overexpression of cyp2j-epoxygenases or sEH regulate KATP channels through A2A AR-cyp2j-PKA-PPAR?/A1 AR-sEH-cyp4a-PPAR¿ pathway in adenosine-induced vascular response. Such results can have a positive impact, as the identified components are expected to provide new targets to curb clinical problems linked with dysfunctional endothelium leading to hypertension.

DESCRIPTION (provided by applicable): Endothelial dysfunction is associated with corresponding changes in vascular tone and increases in contractility, a condition that may cause hypertension in susceptible individuals which may have allelic variants in cyp2j-epoxygenases, solid epoxide hydrolase (sEH), A2A, and A1 adenosine receptors (A2A AR & A1 AR) genes.这些等位基因变体可能与我们的转基因小鼠具有相似之处，后者可能调节血管张力和血压（BP）。我们的初步数据表明，通过A2A AR-CYP2J-PKA-PPAR，腺苷诱导的松弛与开放KATP通道之间可能存在联系？路径。同样，在腺苷诱导的收缩和通过A1 AR-SEH-CYP4A-PPAR途径的KATP通道关闭之间可能存在联系。药物工具和转基因小鼠的结合将使我们能够将可能的靶标确定为治疗可能具有等位基因变体的人群的长期目标。因此，迫切需要探索涉及CYP2J2-氧化酶的可能机制，SEH/CYP4A，A1 AR/A2A AR，PPAR¿/？/？，PKA/PKCCO/？和腺苷诱导的血管反应中的KATP通道。我们的中心假设是，腺苷通过A2A AR-CYP2J-PKA-PPAR通过CYP2J-氧化酶诱导血管松弛并减少BP吗？信号导致打开KATP通道。另一方面，腺苷通过A1 AR-SEH-CYP4A-PPAR途径通过SEH引起血管收缩，并增加BP的增加，导致关闭KATP通道。 To test this hypothesis, we propose to explore in depth mechanism(s) using A2A AR-/-, A1 AR-/-, cyp2j5-/-, sEH-/-, Tie2-cyp2j2Tr (endothelial-cyp2j2 overexpressed), Tie2-sEHTr (endothelial-sEH overexpressed), wild-type mice, immortal renal endothelial cell line from H-2KB-TSA58小鼠，小鼠主动脉内皮细胞（MAEC）和小鼠主动脉平滑肌细胞（MASMC）。此外，我们还将在A2A AR - / - ，CYP2J5 - / - 和TIE2-SEHTR小鼠中使用SEH抑制剂（AUDA/T-AUCB）在饮用水中使用SEH抑制剂（AUDA/T-AUCB）的治疗方法。我们将测量BP，并将使用治疗（腺苷受体激动剂和拮抗剂），CYP-氧化酶，SEH，SEH，Adenyll Cyclase，pkc？ /¿将分析EET和DHET（UPLC-MS/MS）。蛋白质印迹和RT-PCR将用于蛋白质和mRNA表达。我们提出了3个特定目的，以确定：（1）CYP2J- ePxygyass或SEH是否影响BP，腺苷诱导的血管反应和EET/DHET； （2）A2A AR是否存在或不存在通过CYP2J-氧化酶通过PPAR影响腺苷诱导的血管反应，SEH（3）是否存在CYP2J-氧化酶的存在 /过表达或SEH是否通过A2A AR-CYP2A-PKA-PPPAR调节KATP来调节KATP通道ADAR-CYP2J-PKA-PPPAR？血管反应。这样的结果可能会产生积极的影响，因为预计确定的成分将提供新的靶标，以遏制与功能障碍内皮相关的临床问题，导致高血压。