Role of CYP P450s in the Regulation of Vascular Tone through Adenosine Receptors

CYP P450 通过腺苷受体调节血管张力的作用

基本信息

批准号：
8402855
负责人：
S. Jamal Mustafa
金额：
$ 47.1万
依托单位：
WEST VIRGINIA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-01-15 至 2015-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8402855
关键词：
Adenosine Adenosine A1 Receptor Adenosine A2A Receptor Adenosine A3 Receptor Agonist Alkane 1-monooxygenase Aorta Arachidonic Acids Arrhythmia Bathing Blood Vessels Cardiac Cardiomyopathies Cardiovascular Diseases Cardiovascular system Coronary Arteriosclerosis Coupled Cytochromes Data Development Endothelial Cells Endothelium GTP-Binding Proteins Genes Health Heart failure Homeostasis Hydroxyeicosatetraenoic Acids Hypertension Knockout Mice Laboratories Laboratory Study Lead Link Liquid Chromatography MAP Kinase Gene MAP kinase activator MAPK1 gene MAPK3 gene Maintenance Measures Mediating Mediator of activation protein Mitochondria Mitogen-Activated Protein Kinases Mixed Function Oxygenases Monitor Mus Myocardial Infarction Organ Pathway interactions Performance Phosphotransferases Play Population Protein Isoforms Purinergic P1 Receptors Receptor Signaling Regulation Relaxation Role Scheme Signal Pathway Signal Transduction Smooth Muscle Smooth Muscle Myocytes Spectrometry Stroke Surveys Techniques Testing Therapeutic Time Vascular Diseases Vascular Smooth Muscle Vasoconstrictor Agents Vasodilation Western Blotting Work cytochrome P-450 CYP2C subfamily inhibitor/antagonist novel receptor response vasoconstriction

项目摘要

DESCRIPTION (provided by applicant): Adenosine plays an important role in the regulation of vascular tone (VT). The effects of adenosine are mediated by G-protein coupled A1, A2A, A2B and A3 adenosine receptors (AR). This laboratory has made a considerable progress towards the understanding on how adenosine interacts with its receptors to regulate VT. Our earlier studies suggest that vascular smooth muscle and endothelium contain more than one AR subtype that may have opposite actions. The studies from this laboratory demonstrated the role of A2A AR (relaxation) and A1 AR (contraction) in the regulation of VT. Also, this laboratory has demonstrated earlier that endothelium may modulate the actions of adenosine through the release of various mediators. Recently, arachidonic acid (AA)-derived metabolites (EETs & 20-HETE) through CYP450s have emerged as important mediators in cardiovascular system. However, the relationship between AR's and these metabolites in the regulation of VT is not well understood. Therefore, our central hypothesis is that the relaxation caused by A2A AR depends on the activity of CYP epoxygenases while the contraction caused by A1 AR depends on the activity of CYP I-hydroxylases. To test this hypothesis, we will use A2A AR-/-, A2A AR+/+, A1 AR-/- and A1 AR+/+ mice aorta employing organ bath; +/- endothelium; AR agonists & antagonists; CYP epoxygenases inhibitors; EETs & 20-HETE agonists & antagonists; measure EETs and 20-HETE formation through UPLC-MS/MS; signaling in aortic smooth muscle & endothelial cells; Western blot and Real-time PCR techniques. We propose four specific aims to determine whether the presence of A2A or A1 AR can have an effect on the VT through: (a) endothelium by preserving the CYP2C activity to generate EETs (EDHF); (b) smooth muscle CYP4A activity to form 20-HETE; (c) PKC and MAPK (ERK1/ERK2) pathways through CYP4A or CYP2C; and finally, (d) opening or blocking ATP-sensitive (KATP) channel through CYP450s (CYP2C & CYP4A). Findings from these studies can lead to better understanding of the signaling pathways for the regulation of VT by ARs involving endothelium, CYP450s, EETs, 20-HETE, PKC, ERK1/ERK2 kinases and KATP channels. The identification of these pathways for AR signaling and their functional significance in the regulation of VT by adenosine may lead to the development of novel pharmacological agents for the treatment of vascular disorders.

描述（由申请人提供）：腺苷在血管张力调节（VT）中起重要作用。腺苷的作用是由G蛋白偶联的A1，A2A，A2B和A3腺苷受体（AR）介导的。该实验室已经取得了相当大的进步，以理解腺苷如何与受体相互作用以调节VT。我们较早的研究表明，血管平滑肌和内皮包含可能具有相反作用的一种AR亚型。该实验室的研究表明，A2A AR（松弛）和A1 AR（收缩）在VT调节中的作用。此外，该实验室早些时候表明，内皮可以通过释放各种介体来调节腺苷的作用。最近，通过CYP450的蛛网膜酸（AA）衍生的代谢产物（EET和20-HETE）已成为心血管系统中重要的介体。但是，AR和这些代谢物在VT调节中的关系尚不清楚。因此，我们的中心假设是A2A AR引起的弛豫取决于CYP环氧酶的活性，而由A1 AR引起的收缩取决于CYP I-羟基酶的活性。为了检验该假设，我们将使用A2A AR - / - ，A2A AR+/+，A1 AR - / - 和A1 AR+/+小鼠主动脉使用器官浴； +/-内皮； AR激动剂和对手； CYP环氧酶抑制剂； Eets＆20-Hete激动剂和对手；通过UPLC-MS/MS测量EET和20-HETE形成；主动脉平滑肌和内皮细胞中的信号传导；蛋白质印迹和实时PCR技术。我们提出了四个特定的目的，以确定A2A或A1 AR的存在是否可以通过：（a）通过保留CYP2C活性生成EET（EDHF）来对VT产生影响：（a）内皮；（b）平滑肌CYP4A活性形成20-Hete；（c）通过CYP4A或CYP2C的PKC和MAPK（ERK1/ERK2）途径；最后，（d）通过CYP450S（CYP2C和CYP4A）打开或阻止ATP敏感（KATP）通道。这些研究的发现可以更好地理解涉及内皮，CYP450，EET，20-HETE，PKC，ERK1/ERK1/ERK2激酶和KATP通道的ARS调节VT的信号传导途径。这些途径的AR信号传导及其在通过腺苷对VT调节中的功能意义的鉴定可能导致新型药理学剂用于治疗血管疾病。