REDOX REGULATION OF eNOS SIGNALING PATHWAYS IN VASCULAR ENDOTHELIUM

血管内皮细胞 eNOS 信号通路的氧化还原调节

基本信息

批准号：
8250446
负责人：
Thomas Michel
金额：
$ 41.05万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-04-01 至 2015-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8250446
关键词：
5&apos -AMP-activated protein kinase 7,8-dihydrobiopterin AMP-activated protein kinase kinase Affect Biogenesis Biological Availability Biological Markers Biopterin Blood Vessels Calcium Calmodulin Cardiovascular Agents Cardiovascular Diseases Catalysis Caveolae Caveolins Clinical Cysteine Diabetes Mellitus Employee Strikes Endothelial Cells Energy Metabolism Enzyme Inhibition Enzymes Equilibrium Functional disorder Generations Homeostasis Hydrogen Peroxide Hydroxymethylglutaryl-CoA Reductase Inhibitors Instruction Lead Link Mediating Membrane Microdomains Metabolic Metabolism Mitochondria Molecular Mus NADP Nitric Oxide Nitric Oxide Synthase Nitrogen Nitroglycerin Oxidation-Reduction Oxidative Stress Oxidoreductase Oxygen Pathway interactions Phosphorylation Physiology Platelet aggregation Post-Translational Protein Processing Production Protein Isoforms Protein Kinase Protein S Proteins Proto-Oncogene Proteins c-akt RNA Interference Reactive Nitrogen Species Reactive Oxygen Species Recycling Regulation Role Series Signal Pathway Signal Transduction Signaling Protein Small Interfering RNA Sulfhydryl Compounds Superoxides Thioredoxin Vascular Diseases Vascular Endothelial Cell Vascular Endothelium Vasodilation adduct caveolin 1 cofactor glutathione peroxidase human NOS3 protein knock-down oxidation programs protein function research study response scaffold tetrahydrobiopterin

项目摘要

PROJECT SUMMARY (See instructions): The endothelial isoform of nitric oxide synthase (eNOS) is a key determinant of vascular homeostasis and endothelial cell metabolism. eNOS catalysis involves several redox-active cofactors, and NO itself can interact with reactive oxygen species. The bioavailability of endothelium-derived NO is impaired in vascular disease states associated with, increased oxidative stress. We hypothesize that redox regulation ofthe eNOS pathway represents a critical determinant of NO- and ROS-dependent signaling and metabolic regulation in vascular endothelial cells. The proposed studies have important points of intersection with experimental plans proposed by other Projects in this Program. eNOS catalysis involves several redox-active cofactors; changes in intracellular redox state affect the concentrations of these key cofactors, and lead to alterations in eNOS-modulated responses. The formation of cellular NO adducts is influenced by reactive nitrogen and reactive oxygen species and by the cellular thiol redox state. Phosphorylation of eNOS by the AMP-activated protein kinase (AMPK) is influenced by reactive oxygen species, providing an important link between oxidative stress, eNOS signaling, and endothelial cell metabolism. Statins also activate AMPK and modulate endothelial ROS production. eNOSJs targeted to signal-transducing membrane microdomains termed caveolae, where the enzyme interacts with the scaffolding/signaling protein caveolin-1. We discovered that siRNA-mediated knockdown of caveolin leads to a striking increase in ROS production from endothelial cells, and found that caveolin-1-/- mice show dramatic increases in oxidative stress. eNOS-caveolin interactions are modulated by statins, yet the roles of statins in modulation of redox pathways involving eNOS remain incompletely understood. Experiments proposed in Specific Aim 1 will identify the mechanisms whereby altered biopterin and thiol metabolism affects eNOS post-translational modifications, subcellular targeting, and ROS generation. Experiments in Specific Aim 2 will determine the mechanisms whereby caveolin, statins, and reactive oxygen species modulate AMP-activated protein kinase (AMPK) signaling to eNOS.

项目摘要（请参阅说明）：一氧化氮合酶（ENOS）的内皮同工型是血管稳态的关键决定因素，内皮细胞代谢。 eNOS催化涉及几个氧化还原活性辅助因子，没有任何人可以与活性氧相互作用。内皮衍生的NO的生物利用度在血管中受损与氧化应激相关的疾病状态。我们假设对eNOS的氧化还原调节途径代表了不依赖和ROS的信号传导和代谢调节的关键决定因素血管内皮细胞。提出的研究具有与实验的相交的重要点该计划中其他项目提出的计划。 eNOS催化涉及几个氧化还原活性辅助因子。细胞内氧化还原状态的变化会影响这些关键辅助因子的浓度，并导致改变 eNOS调节的响应。细胞的形成无内合物受反应性氮的影响和活性氧和通过细胞硫醇氧化还原状态。通过AMP激活的ENOS磷酸化蛋白激酶（AMPK）受活性氧的影响，提供了重要的联系氧化应激，eNOS信号传导和内皮细胞代谢。他汀类药物还激活AMPK并调节内皮ROS产生。针对信号传递的膜微域的eNOSJS 小窝，酶与脚手架/信号蛋白小窝蛋白1相互作用。我们发现了这一点 siRNA介导的小窝蛋白敲低导致内皮细胞ROS产生的显着增加，并发现小窝蛋白1 - / - 小鼠在氧化应激中显示出急剧增加。 eNOS-caveolin相互作用由他汀类药物调节，但他汀类药物在涉及eNOS的氧化还原途径调节中的作用仍然存在不完全理解。特定目标1中提出的实验将确定该机制生物肽和硫醇代谢改变会影响eNOS翻译后修饰，亚细胞靶向，和Ros Generation。特定AIM 2中的实验将确定可窝蛋白的机制，他汀类药物和活性氧物种调节AMP激活的蛋白激酶（AMPK）信号传导到eNOS。