Reactive Oxygen Species and Vascular O2 Sensing

活性氧和血管 O2 传感

• 批准号: 7330318
• 负责人: Michael S Wolin
• 金额: $ 30.3万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-30 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7330318
• 关键词: Arteries; Blood Vessels; Bos taurus; Calcium; Cattle; Coronary; Coronary artery; Cyclic GMP; Depth; Detection; Endothelium; Enzymes; Extracellular Signal Regulated Kinases; Fluorescence; Generations; Glucosephosphate Dehydrogenase; Glutathione; Hand; High Pressure Liquid Chromatography; Hydrogen Peroxide; Hypoxia; Investigation; Laboratories; Link; Lung; Measurement; Mediating; Metabolic; Metabolic Activation; Microscopic; Muscle; NADH; NADP; Organoids; Oxidants; Oxidases; Oxidation-Reduction; Pathway interactions; Pentosephosphate Pathway; Peroxides; Physiological; Preparation; Process; Production; Property; Pulmonary artery structure; Rattus; Reactive Oxygen Species; Regulation; Relaxation; Resistance; Role; Signal Transduction; Small Interfering RNA; Smooth Muscle Myocytes; Soluble Guanylate Cyclase; Source; Superoxides; System; Techniques; Tissues; Transfection; Vascular Smooth Muscle; Work; cellular imaging; design; enzyme activity; fluorescence imaging; inorganic phosphate; oxidation; response

This proposal focuses on defining the role of Nox oxidase-derived hydrogen peroxide, and cytosolic NADPH and NADH redox systems in processes that control signaling mechanisms regulating vascular smooth muscle force responses to physiological changes in PO2.Studies in this application investigate the importance of our new evidence that cytosolic NADPH and possibly NADH levels control a baseline Nox oxidase-derived hydrogen peroxide-mediated relaxation of bovine pulmdnary arteries which is removed by hypoxia, resulting in contraction. On the other hand, bovine coronary arteries maintain a lower level of NADPH and the NADPH generating pentose phosphate pathway (PPP) enzyme glucose-6-phosphate dehydrogenase. We hypothesize that differences in the function of the PPP in coronary arteries results in a hypoxia-elicited oxidation of cytosolic NADPH, which activates relaxation through a mechanism we have shown to be controlled by the PPP.Studies in Aim 1 will investigate how PO2 regulates the expression of vascular responses involving Nox-derived hydrogen peroxide. Aim 2 focuses on understanding how PO2 regulates cytosolic NADH and NADPH redox-linked signaling mechanisms that contribute to the control of force generation. Studies in Aim 3 are to define the origins of differences in the control of Nox oxidase activity and cytosolic NAD(P)H redox that contribute to hypoxia-elicited contractile and relaxing responses observed in pulmonary and coronary arteries. Isolated endothelium-removed coronary and pulmonary conduit and resistance arteries will be studied with perturbations that alter Nox activity and expression, and cytosolic NADH and NADPH redox to define how signaling systems regulated by these processes may function in PO2-elicited responses. The control of cytosolic NAD(P)H redox and oxidant production by PO2 will be examined with a combination of metabolic measurements, tissue fluorescence and chemiluminescent techniques, and cellular fluorescence imaging approaches which are designed to characterize the ROS and redox aspects of the signaling mechanisms that are involved. Rat arteries will also be examined to identify similarities and differences in mechanisms of PO2-elicited responses controlled by Nox and cytosolic NAD(P)H redox. These studies should help define the origins of differences in physiological responses of pulmonary and coronary arteries to changes in O2 tension.

这项建议的重点是确定氮氧化物氧化酶衍生的过氧化氢和细胞溶质NADPH的作用， 和NADH氧化还原系统在控制调节血管平滑肌的信号传导机制的过程中的作用 肌肉力量对PO 2生理变化的反应。 我们的新证据的重要性，胞质NADPH和可能的NADH水平控制基线氮氧化物 氧化酶衍生的过氧化氢介导的牛肺动脉舒张， 缺氧导致收缩另一方面，牛冠状动脉维持较低水平的 NADPH和NADPH生成戊糖磷酸途径（PPP）酶葡萄糖-6-磷酸 脱氢酶。我们假设，在冠状动脉PPP功能的差异，导致在一个 缺氧引起的细胞质NADPH氧化，通过我们已知的机制激活松弛 目的1中的研究将研究PO 2如何调节 血管反应涉及氮氧化物衍生的过氧化氢。目标2：了解PO 2 调节胞质NADH和NADPH氧化还原相关的信号传导机制，有助于控制 力量生成目标3中的研究是确定Nox氧化酶控制差异的起源 活性和胞质NAD（P）H氧化还原，有助于缺氧引起的收缩和舒张反应 在肺动脉和冠状动脉中观察到。去内皮的冠状动脉和肺动脉 导管和阻力动脉将被研究与扰动，改变氮氧化物的活性和表达， 胞质NADH和NADPH氧化还原，以确定如何信号系统调节这些过程可能 在PO 2引起的反应中起作用。PO 2对胞质NAD（P）H氧化还原和氧化剂产生的调控 将结合代谢测量、组织荧光和荧光发光进行检查 技术和细胞荧光成像方法，其被设计用于表征ROS和 涉及的信号机制的氧化还原方面。还将检查大鼠动脉， NO_x和胞浆NO_x控制的PO_2诱导反应机制的异同 NAD（P）H氧化还原。这些研究应该有助于确定不同的生理反应的起源， 肺动脉和冠状动脉的氧分压变化。