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

• 批准号: 7564164
• 负责人: S. Jamal Mustafa
• 金额: $ 50.32万
• 依托单位: WEST VIRGINIA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-01-15 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7564164
• 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; 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; public health relevance; receptor; response; vasoconstriction

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. PUBLIC HEALTH RELEVANCE: Adenosine plays an important role in the maintenance of vascular tone via activation of four receptor (AR) subtypes. In blood vessels, vasodilation is primarily caused by the activation of A2A. Survey of cytochrome 450s (CYP) expression among populations have shown that there is a link between CYP expression and cardiovascular disease, such as hypertension, coronary artery disease, myocardial infarction, heart failure, stroke, cardiomyopathy and arrhythmias. Evidence is mounting for the role of CYP metabolites (EETs or DHETs or 20HETE) in the maintenance of cardiovascular health, including the regulation of vascular tone and cardiac contractility. This proposal is directed towards understanding the relationship between ARs and CYP activity and the relationship between ARs and CYP-generated metabolites (EETs or DHETs or 20HETE). The data generated from this project may lead to better therapeutic approaches involving the imbalance and dysregulation in CYP metabolite(s) formation in cardiovascular diseases.

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。 公共卫生相关性：通过激活四种受体（AR）亚型维持血管张力，腺苷在维持血管张力中起着重要作用。在血管中，血管舒张主要是由A2a的激活引起的。人群中细胞色素450（CYP）表达的调查表明，CYP表达与心血管疾病之间存在联系，例如高血压，冠状动脉疾病，心肌梗死，心力衰竭，中风，心肌病和心律失常。有证据表明CYP代谢物（EET或DHET或20HETE）在维持心血管健康中的作用，包括调节血管张力和心脏收缩率。该建议是针对理解ARS与CYP活动之间的关系，以及ARS与CYP生成的代谢产物（EET或DHET或20HETE）之间的关系。该项目产生的数据可能导致更好的治疗方法，涉及心血管疾病中CYP代谢产物形成的不平衡和失调。