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（申请人提供）：腺苷在调节血管张力（VT）中发挥重要作用。腺苷的作用由 G 蛋白偶联的 A1、A2A、A2B 和 A3 腺苷受体 (AR) 介导。该实验室在了解腺苷如何与其受体相互作用以调节 VT 方面取得了相当大的进展。我们早期的研究表明，血管平滑肌和内皮细胞含有不止一种 AR 亚型，这些亚型可能具有相反的作用。该实验室的研究证明了 A2A AR（放松）和 A1 AR（收缩）在 VT 调节中的作用。此外，该实验室早些时候已经证明内皮细胞可以通过释放各种介质来调节腺苷的作用。最近，花生四烯酸（AA）衍生的代谢物（EET 和 20-HETE）通过 CYP450 已成为心血管系统中的重要介质。然而，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 引起的松弛取决于 CYP 环氧化酶的活性，而 A1 AR 引起的收缩取决于 CYP I-羟化酶的活性。为了验证这一假设，我们将使用 A2A AR-/-、A2A AR+/+、A1 AR-/- 和 A1 AR+/+ 小鼠主动脉进行器官浴； +/-内皮； AR激动剂和拮抗剂； CYP环氧化酶抑制剂； EET 和 20-HETE 激动剂和拮抗剂；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。我们提出了四个具体目标，以确定 A2A 或 A1 AR 的存在是否可以通过以下方式对 VT 产生影响： (a) 内皮细胞通过保留 CYP2C 活性来产生 EET (EDHF)； (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 CYP450（CYP2C 和 CYP4A）打开或阻断 ATP 敏感 (KATP) 通道。这些研究的结果可以帮助我们更好地了解 AR 调节 VT 的信号通路，涉及内皮、CYP450、EET、20-HETE、PKC、ERK1/ERK2 激酶和 KATP 通道。这些 AR 信号通路的鉴定及其在腺苷调节 VT 中的功能意义可能会导致治疗血管疾病的新型药物的开发。 公共健康相关性：腺苷通过激活四种受体 (AR) 亚型在维持血管张力方面发挥重要作用。在血管中，血管舒张主要是由 A2A 的激活引起的。对人群中细胞色素450s（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、心肌梗塞、心力衰竭、中风、心肌病和心律失常。越来越多的证据表明 CYP 代谢物（EET 或 DHET 或 20HETE）在维持心血管健康（包括调节血管张力和心肌收缩力）中的作用。该提案旨在了解 AR 与 CYP 活性之间的关系以及 AR 与 CYP 生成的代谢物（EET 或 DHET 或 20HETE）之间的关系。该项目产生的数据可能会带来更好的治疗方法，涉及心血管疾病中 CYP 代谢物形成的不平衡和失调。