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

基本信息

批准号：
8666036
负责人：
Mohammed A Nayeem
金额：
$ 36.97万
依托单位：
WEST VIRGINIA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-06-01 至 2015-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8666036
关键词：
ADORA2A gene Acetylcholine Adenosine Adenosine A1 Receptor Adenylate Cyclase Affect Agonist Antiinflammatory Effect 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 GTP-Binding Proteins Genes Genetic Polymorphism Goals Health Human Hypertension Image Individual Kidney Link Measures Mediating Mitogen-Activated Protein Kinase Inhibitor Mus Myocardial perfusion Organ Outcome PKA inhibitor PPAR gamma Pathway interactions Patients Pharmaceutical Preparations Pharmacology Population Prevention Protein Kinase C Alpha 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 endothelial dysfunction genetic variant inhibitor/antagonist innovation knockout gene mRNA Expression novel overexpression renal artery response tool vasoconstriction

项目摘要

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-PPARγ pathway. Also, it is possible that a link may exist between adenosine-induced contraction and closing of KATP channels through A1 AR-sEH-cyp4a- PPARα 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, PPARα/γ, PKA/PKCα/ϵ 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-PPARγ 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-PPARα 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, PKCα/ϵ, MAPK and PKA inhibitors, PPARα/γ, 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-PPARγ/A1 AR-sEH-cyp4a-PPARα 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.

描述（由申请人提供）：内皮功能障碍与血管张力的相应变化和收缩力增加有关，这种情况可能导致易感个体高血压，这些个体可能具有 cyp2j-环氧合酶、可溶性环氧化物水解酶 (sEH)、A2A 等位基因变异，和 A1 腺苷受体（A2A AR 和 A1 AR）基因这些等位基因变体可能与我们的转基因小鼠有相似之处，可以调节。我们的初步数据表明，腺苷通过 A2A AR-cyp2j-PKA-PPARγ 途径诱导的松弛和 KATP 通道开放之间可能存在联系。 -通过 A1 AR-sEH-cyp4a-PPARα 途径诱导 KATP 通道收缩和关闭药理学工具和转基因小鼠的结合将使我们能够确定可能的长期目标。因此，迫切需要探索涉及 cyp2j2-环氧合酶、sEH/cyp4a、A1 AR/A2A AR、PPARα/γ、PKA/PKCα/ϵ 和 KATP 的可能机制。我们的中心假设是腺苷通过诱导血管舒张和血压降低。 cyp2j-环氧合酶通过 A2A AR-cyp2j-PKA-PPARγ 信号传导导致 KATP 通道打开，另一方面，腺苷通过 sEH 通过 A1 AR-sEH-cyp4a-PPARα 途径诱导血管收缩和血压升高，导致 KATP 关闭。为了检验这一假设，我们建议使用 A2A AR-/-、A1 深入探索机制。 AR-/-、cyp2j5-/-、sEH-/-、Tie2-cyp2j2Tr（内皮-cyp2j2 过表达）、Tie2-sEHTr（内皮-sEH 过表达）、野生型小鼠、来自 H-2Kb- 的永生肾内皮细胞系tsA58 小鼠、小鼠主动脉内皮细胞 (MAEC) 和小鼠主动脉平滑肌细胞此外，我们还将探索在饮用水（或灌胃）中使用 sEH 抑制剂 (AUDA/t-AUCB) 对 A2A AR-/-、cyp2j5-/- 和 Tie2-sEHTr 小鼠进行治疗的可能性。血压高。我们将测量血压，并使用主动脉/肾动脉（器官浴/DMT 线肌动描记器）进行治疗（腺苷受体）激动剂和拮抗剂）、cyp-环氧合酶、sEH、腺苷酸环化酶、PKCα/ϵ、MAPK 和 PKA 抑制剂、PPARα/γ、EET 和 KATP 通道（激活剂和抑制剂）将用于蛋白质和 mRNA 的蛋白质印迹和 RT-PCR。我们提出3个具体目标来确定：（1）是否。 cyp2j-环氧合酶或 sEH 影响血压、腺苷诱导的血管反应和 EET/DHET；(2) A2A AR 的存在或不存在是否通过 cyp2j-环氧合酶、sEH 影响腺苷诱导的血管反应；(3) A2A AR 是否存在/cyp2j-环氧合酶或 sEH 的过度表达通过以下方式调节 KATP 通道A2A AR-cyp2j-PKA-PPARγ/A1 AR-sEH-cyp4a-PPARα 通路在腺苷诱导的血管反应中可以产生积极的影响，因为所确定的成分有望提供新的靶点来遏制与功能失调相关的临床问题。内皮细胞导致高血压。