Thioredoxin and Endothelial Cell Function

硫氧还蛋白和内皮细胞功能

• 批准号: 7896738
• 负责人: WANG MIN
• 金额: $ 41.38万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7896738
• 关键词: Antioxidants; Aorta; Apolipoprotein E; Apoptosis; Apoptotic; Arterial Fatty Streak; Arteries; Atherosclerosis; Binding; Biochemistry; Biological Availability; Blood Vessels; Blood flow; Breeding; Catalytic Domain; Cause of Death; Cell physiology; Cells; Cessation of life; Complex; Coronary artery; Data; Development; Embryo; Endothelial Cells; Endothelium; Environment; Enzymes; Figs - dietary; Fluorescence Resonance Energy Transfer; Fractionation; Functional disorder; Generations; Genetic; Hydrogen Peroxide; In Vitro; Inflammation; Inflammatory; Knock-out; Knockout Mice; Lead; MAP3K5 gene; Mediating; Mediator of activation protein; Mitochondria; Mus; Myocardial Infarction; Organelles; Oxidases; Oxidation-Reduction; Oxidative Stress; Oxidoreductase; Pathway interactions; Patients; Peroxidases; Plasma; Play; Process; Production; Proteins; Reactive Oxygen Species; Regulation; Role; Signal Transduction; Stimulus; Stress; Superoxide Dismutase; Superoxides; System; TNF gene; Testing; Thioredoxin; Transgenic Mice; Transgenic Organisms; United States; Vascular Endothelium; Vasodilation; Viral Vector; Work; base; catalase; cellular targeting; cytokine; improved; in vivo; in vivo Model; mouse model; mutant; novel therapeutic intervention; overexpression; oxidation; prevent; protective effect; protein protein interaction; response

DESCRIPTION (provided by applicant): Myocardial infarction due to atherosclerosis of coronary arteries remains the leading cause of death in the United States. It has become clear that changes of cellular/systemic redox state, resulting in increases in inflammation (e.g., TNF) and reactive oxygen species (ROS), represent a common pathogenic mechanism for atherosclerosis. The vascular cell that primarily limits the inflammatory and atherosclerotic process is the endothelial cell (EC). ROS-induced reduction in NO bioavailability and increase of EC apoptosis results in a proatherogenic state. Increasing evidence supports that ROS generated from mitochondria in vasculature significantly contribute to EC dysfunction and atherosclerotic progression. Furthermore, recent data suggest that mitochondria normally produce the strongest reducing environment among all cellular organelles, and mitochondria are especially vulnerable to oxidation in response to stress stimuli including proinflammatory cytokines. A key system regulating mitochondria redox is mitochondria-specific thioredoxin (Trx2) system, consisting of Trx2, Trx2 reductase (TrxR2) and Trx2-depndent peroxidase (Prx3). Little is known for the role of mitochondrial Trx2 system in vasculature. Our data suggest that mitochondrial Trx2 may play critical roles in maintaining mitochondria reduced state, preventing ROS-induced EC dysfunction. Specifically, we have used both EC-specific transgenic and knockout mice, and demonstrated a critical role of Trx2 in regulating endothelium functions by increasing NO bioactivity. We also show that Trx2 inhibits the activity of proapoptotic protein kinase ASK1 through protein-protein interactions, protecting EC from TNF and ROS- induced apoptosis. We propose that Trx2 prevents ROS-induced EC dysfunction through two distinct and cooperative pathways: Trx2 maintains a reduced state of mitochondria in EC, reducing ROS generation leading and increasing NO bioactivity; Trx2 protects ROS-induced EC apoptosis by directly binding to ASK1. We further hypothesize that increased NO bioactivity and decreased apoptotic responses prevent EC dysfunction and atherosclerotic development. To explore these hypotheses, we propose the following specific aims: 1) Determine the mechanisms by which Trx2 preserves NO bioactivity and EC function. 2) Determine the mechanisms by which Trx2 inhibits mitochondrial ASK1-mediated apoptosis. 3) Determine the role of Trx2 in preventing EC dysfunction and atherosclerosis development/progression in a mouse model. This proposal uses both in vitro and in vivo models to determine the roles of Trx2 in protection against ROS- induced EC dysfunction and atherosclerosis development/progression. These studies, if successful, will facilitate the development of new therapeutic approaches to control atherosclerosis progression and myocardial infarction. Project Narrative: Myocardial infarction due to narrowing of arteries manifesting as decreased blood flow remains the leading cause of death in the United States. We will study the effects of a antioxidant protein thioredoxin, on vascular endothelium. Our work may lead to better tests and treatments for atherosclerosis patients.

描述（由申请人提供）：冠状动脉粥样硬化引起的心肌梗死仍然是美国死亡的主要原因。很明显，细胞/全身氧化还原状态的改变，导致炎症（如TNF）和活性氧（ROS）的增加，是动脉粥样硬化的常见致病机制。主要限制炎症和动脉粥样硬化过程的血管细胞是内皮细胞（EC）。ros诱导的NO生物利用度降低和EC细胞凋亡增加导致动脉粥样硬化前期状态。越来越多的证据支持血管中线粒体产生的活性氧显著促进EC功能障碍和动脉粥样硬化进展。此外，最近的数据表明，线粒体通常在所有细胞器中产生最强的还原环境，并且线粒体在应对包括促炎细胞因子在内的应激刺激时特别容易氧化。调控线粒体氧化还原的关键系统是线粒体特异性硫氧还蛋白（Trx2）系统，由Trx2、Trx2还原酶（TrxR2）和Trx2依赖性过氧化物酶（Prx3）组成。线粒体Trx2系统在血管系统中的作用知之甚少。我们的数据表明，线粒体Trx2可能在维持线粒体减少状态，防止ros诱导的EC功能障碍中发挥关键作用。具体来说，我们使用了ec特异性转基因和敲除小鼠，并证明了Trx2通过增加NO生物活性在调节内皮功能方面的关键作用。我们还发现Trx2通过蛋白-蛋白相互作用抑制促凋亡蛋白激酶ASK1的活性，保护EC免受TNF和ROS诱导的凋亡。我们认为Trx2通过两种不同的合作途径阻止ROS诱导的EC功能障碍：Trx2维持EC中线粒体的减少状态，减少ROS的产生并增加NO的生物活性；Trx2通过直接结合ASK1保护ros诱导的EC凋亡。我们进一步假设，一氧化氮生物活性的增加和凋亡反应的减少可以防止EC功能障碍和动脉粥样硬化的发展。为了探索这些假设，我们提出以下具体目标：1)确定Trx2保持NO生物活性和EC功能的机制。2)确定Trx2抑制线粒体ask1介导的细胞凋亡的机制。3)在小鼠模型中确定Trx2在预防EC功能障碍和动脉粥样硬化发生/进展中的作用。本研究采用体外和体内模型来确定Trx2在防止ROS诱导的EC功能障碍和动脉粥样硬化发生/进展中的作用。这些研究如果成功，将促进新的治疗方法的发展，以控制动脉粥样硬化的进展和心肌梗死。项目描述：动脉狭窄引起的心肌梗死表现为血流减少，在美国仍然是导致死亡的主要原因。我们将研究一种抗氧化蛋白硫氧还蛋白对血管内皮的影响。我们的工作可能会为动脉粥样硬化患者带来更好的测试和治疗。