Redox Regulation of Endothelial Nitric Oxide Synthase and Cardiovascular Diseases

项目摘要： 这个职业发展奖的目的是继续发展春安博士的学术生涯 陈博士最初是俄亥俄州州立大学心肺研究所的博士后研究员， 额外的博士后培训和过渡到心血管领域的独立研究者 专门从事NOS功能和氧化还原信号的研究。增加氧自由基的产生， 可降低一氧化氮的生物利用度，被认为是缺血/再灌注的主要发病机制 受伤在内皮细胞中，内皮型一氧化氮合酶（eNOS）是产生一氧化氮合酶的重要酶。 这个维持心血管功能的关键分子。越来越多的证据表明， 氧化应激通过氧化翻译后修饰改变了几种酶的功能，例如 如S-谷胱甘肽化、硝化或亚硝基化，所有这些都与信号转导有关。 在辅导阶段，目标1和目标2将解决几个问题：目标1是确定 eNOS S-谷胱甘肽化和巯基氧化的详细机制，因为它与心血管疾病有关。 目的2是确定Grx 1在脱谷胱甘肽化过程中的作用，氧化还原调节和NOS功能。 从指导阶段培训中获得的结果将为指导阶段和 独立阶段。在独立阶段，陈博士的研究将集中在离体小鼠模型上 关于eNOS S-谷胱甘肽化对血管功能的影响，以及Grx 1在 缺血/再灌注损伤几个机械问题将解决以下目标：目标3是 研究eNOS Cys突变体对血管功能和抗氧化修饰的作用， 血管功能障碍目的4是确定Grx 1在缺血/再灌注损伤中的作用，特别是在缺血/再灌注损伤中的作用。 调节蛋白脱谷胱甘肽和eNOS功能，在小鼠离体心脏模型。通过了解 氧化还原调节酶（Grx）与NO产生之间关系，这将提供一个关键步骤 为了了解在心血管功能的改变过程中所涉及的机制， 缺血/再灌注和氧化应激。 成功获得K99/R 00奖将促进陈春安博士的职业研究发展， 使Chun-An Chen博士成为心血管研究领域的独立研究者。

Project Summary: The objective of this career development award is to continue to develop the academic career of Dr. Chun-An Chen, first as a postdoctoral researcher at The Ohio State University Heart and Lung Research Institute, with additional postdoctoral training and transition to an independent investigator in the field of cardiovascular research specializing in NOS function and redox signaling. Increased oxygen free radical generation, which can reduce the bioavailability of nitric oxide, is believed to be the primary pathogenesis of ischemia/reperfusion injuries. In the endothelium, endothelial nitric oxide synthase (eNOS) is the important enzyme that produces this critical molecule maintaining the cardiovascular function. Growing evidence suggests that increased oxidative stress alters the function of several enzymes through oxidative post-translational modifications, such as S-glutathionylation, nitration, or nitrosylation, all of which have been implicated in signal transduction. During the mentored phase, several questions will be addressed in Aim1 and Aim2: Aim1 is to determine the detailed mechanism of eNOS S-glutathionylation and thiol oxidation, as it pertains to cardiovascular diseases. Aim2 is to determine the role of Grx1 in the deglutathionylation process, redox regulation, and NOS function. The results gained from the mentored phase training will provide a bridge between mentored phase and independent phase. During the independent phase, Dr. Chen's research will focus on ex vivo mouse models regarding the effect of eNOS S-glutathionylation on vascular function, and the role of Grx1 on ischemia/reperfusion injury. Several mechanistic questions will be addressed with the following aims: Aim3 is to study the effect of eNOS Cys mutants on vascular function and resistance to oxidative modification during vascular dysfunction. Aim4 is to identify the role of Grx1 in ischemia/reperfusion injury, especially in the regulation of protein deglutathionylation and eNOS function, in mouse ex vivo heart models. By understanding the relationship between a redox regulatory enzyme (Grx) and NO production, this will provide a critical step toward understanding the mechanisms involved in the alteration of cardiovascular function during ischemia/reperfusion and oxidative stress. Successfully securing this K99/R00 award will foster Dr. Chun-An Chen's career research development and enable Dr. Chun-An Chen to become an independent investigator in the field of cardiovascular research.