Superoxide Generation from eNOS: The Role of Pterins

eNOS 产生超氧化物：蝶呤的作用

• 批准号: 7894501
• 负责人: JEANNETTE M. VASQUEZ VIVAR
• 金额: $ 26.51万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-12-16 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7894501
• 关键词: 4 hydroxynonenal; Animal Model; Anions; Antioxidants; Arterial Fatty Streak; Atherosclerosis; Biopterin; Blood Vessels; Calcium; Cardiovascular Diseases; Caspase; Cell Adhesion; Cell Adhesion Molecules; Cell Culture Techniques; Cell Death; Cell physiology; Cells; Chelating Agents; Cholesterol; Complex; Deet; Detection; Disease; Endothelial Cells; Endothelium; Esters; Etiology; Evaluation; Event; Experimental Models; Figs - dietary; Flow Cytometry; Fluorescence; Functional disorder; Funding; GTP Cyclohydrolase I; Generations; Goals; High Pressure Liquid Chromatography; Human; Knowledge; Laboratories; Lead; Link; Lipid Peroxidation; Lipid Peroxides; Lipids; Mediating; Mediator of activation protein; Membrane Potentials; Metabolic Pathway; Metabolism; Mitochondria; Nitric Oxide; Nitric Oxide Donors; Nitric Oxide Synthase; Oxidants; Oxidation-Reduction; Oxidative Stress; Oxygen; Pathology; Peroxides; Pharmaceutical Preparations; Phenotype; Physiological; Physiology; Play; Prevention; Production; Property; Proteasome Inhibitor; Proteins; Pterins; Reactive Oxygen Species; Recycling; Regulation; Relaxation; Research Personnel; Risk Factors; Role; Signal Transduction; Source; Sulfhydryl Compounds; Superoxides; Supplementation; Testing; Transgenic Mice; Variant; antioxidant therapy; atherogenesis; base; cofactor; design; genetic manipulation; human NOS3 protein; improved; inhibitor/antagonist; loss of function; mitochondrial dysfunction; mitochondrial membrane; multicatalytic endopeptidase complex; outcome forecast; overexpression; oxidation; oxidative damage; peroxidation; programs; research study; response; sepiapterin; tetrahydrobiopterin; vasoconstriction

DESCRIPTION (provided by applicant): Endothelial dysfunction (ED) is a critical event in the pathophysiology of atherosclerosis and other cardiovascular diseases (CVD). The underlying causes of ED have not been fully established, although unbalanced production of nitric oxide (NO) and reactive oxygen production (ROS) are causally involved. Enzymatic production of NO is dependent on optimal basal tetrahydrobiopterin (BH4) levels in the endothelium. Increased availability of the cofactor by genetic manipulation or pharmacological supplementation has been shown to be beneficial, although the mechanisms controlling BH4 in the endothelium are poorly understood. The broad objectives of this renewal is designed to bridge the gap in knowledge and is based upon the hypothesis that altering BH4 metabolism by lipid peroxidation products and ROS has important consequences in normal NO/ROS fluxes and endothelial physiology favoring phenotypical changes associated with atherogenesis. The present proposal is built on three findings: (i) BH4-free endothelial nitric oxide synthase (eNOS) generates ~100 nmoles superoxide/min/mg protein which is inhibited by BH4 (micromolar range); (ii) BH4 depletion in endothelial cells increases superoxide production by calcium-dependent mechanisms; (iii) 4-hydroxy-2-nonenal (HNE) is a potent inhibitor of BH4 synthesis, depletes BH4 and NO, and increases superoxide production in endothelial cells. The evaluation of the consequences of increased superoxide production from eNOS is critical to the implications of NO and ROS oxidant signaling in disease. Specifically we will: 1) establish the mechanisms increasing uncoupled eNOS-dependent superoxide by HNE and peroxides; 2) investigate the influence of BH4 depletion by HNE and peroxides on increased mitochondrial-superoxide release and dysfunction; 3) elucidate the role of BH4 in limiting oxidative damage, mitochondrial dysfunction and changes in cell phenotype. In the execution of this proposal we will use established endothelial cell cultures and also cells isolated from GTPCH-transgenic mice to examine the influence of endogenous variation in BH4 in the endothelial responses which have been linked to protection, "re-coupling" of eNOS and/or additional antioxidant activity. Generally this proposal is aimed at understanding these fundamental mechanisms that should provide the basis for developing new and improved strategies in the prevention and treatment of atherosclerosis and CVD.

描述(申请人提供)：内皮功能障碍(ED)是动脉粥样硬化和其他心血管疾病(CVD)病理生理学中的关键事件。ED的根本原因尚未完全确定，尽管一氧化氮(NO)和反应性氧(ROS)的不平衡产生是原因之一。酶促NO的产生依赖于内皮细胞中最适的基础四氢生物蝶呤(BH4)水平。通过基因操作或药物补充增加辅因子的可用性已被证明是有益的，尽管控制内皮细胞中BH4的机制尚不清楚。这一更新的广泛目标旨在弥合知识的差距，并基于这样的假设，即通过脂质过氧化产物和ROS改变BH4代谢对正常的NO/ROS通量和有利于与动脉粥样硬化相关的表型变化的内皮生理学具有重要影响。目前的建议基于三个发现：(I)无BH4的内皮型一氧化氮合酶(ENOS)产生约100nmoles的超氧化物/分钟/毫克蛋白，BH4(微摩尔范围)抑制该蛋白；(Ii)内皮细胞中BH4的耗竭通过钙依赖机制增加超氧化物的产生；(Iii)4-羟基-2-壬烯醛(HNE)是一种有效的BH4合成抑制剂，可消耗BH4和NO，并增加内皮细胞中超氧化物的产生。对eNOS产生的超氧化物增加的后果的评估对于NO和ROS氧化剂信号在疾病中的意义至关重要。具体地说，我们将：1)建立HNE和过氧化产物增加解偶联eNOS依赖的超氧化物的机制；2)研究HNE和过氧化产物耗尽BH4对增加线粒体超氧化物释放和功能障碍的影响；3)阐明BH4在限制氧化损伤、线粒体功能障碍和细胞表型变化中的作用。在这项建议的执行过程中，我们将使用已建立的内皮细胞培养和从GTPCH转基因小鼠分离的细胞来检测BH4内源性变异对内皮反应的影响，这些反应与保护、eNOS的“重新偶联”和/或额外的抗氧化活性有关。总的来说，这项建议旨在了解这些基本机制，为制定预防和治疗动脉粥样硬化和心血管疾病的新的和改进的战略奠定基础。

项目成果

Pulmonary hypertension in the newborn GTP cyclohydrolase I-deficient mouse.

• DOI: 10.1016/j.freeradbiomed.2011.09.012
• 发表时间: 2011-12-15
• 期刊: FREE RADICAL BIOLOGY AND MEDICINE
• 影响因子: 7.4
• 作者: Belik, Jaques;McIntyre, Brendan A. S.;Enomoto, Masahiro;Pan, Jingyi;Grasemann, Hartmut;Vasquez-Vivar, Jeannette
• 通讯作者: Vasquez-Vivar, Jeannette

NAD(P)H oxidase and eNOS play differential roles in cytomegalovirus infection-induced microvascular dysfunction.

• DOI: 10.1016/j.freeradbiomed.2011.09.039
• 发表时间: 2011-12-15
• 期刊: FREE RADICAL BIOLOGY AND MEDICINE
• 影响因子: 7.4
• 作者: Leskov, Igor L.;Whitsett, Jennifer;Vasquez-Vivar, Jeannette;Stokes, Karen Y.
• 通讯作者: Stokes, Karen Y.

Human endothelial dihydrofolate reductase low activity limits vascular tetrahydrobiopterin recycling.

• DOI: 10.1016/j.freeradbiomed.2013.04.035
• 发表时间: 2013-10
• 期刊: FREE RADICAL BIOLOGY AND MEDICINE
• 影响因子: 7.4
• 作者: Whitsett, Jennifer;Range Filho, Artur;Sethumadhavan, Savitha;Celinska, Joanna;Widlansky, Michael;Vasquez-Vivar, Jeannette
• 通讯作者: Vasquez-Vivar, Jeannette