Hsp90, NOS3 and Cardioprotection

Hsp90、NOS3 和心脏保护

• 批准号: 7332187
• 负责人: Yang Shi
• 金额: $ 36.78万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7332187
• 关键词: Anabolism; Biopterin; Cardiovascular Physiology; Cessation of life; Count; Coupled; Data; Development; Diabetes Mellitus; Diet; Direct Costs; Enzymes; Equilibrium; Exhibits; GTP Cyclohydrolase I; Generations; Goals; Healthcare; Heart; Heart Injuries; Heat-Shock Proteins 90; Human; Inbred Strain; Inbred Strains Rats; Injury; Insulin Resistance; Ischemia; Lead; Measurement; Mediating; Modeling; Molecular; Molecular Chaperones; Myocardial; Myocardial Ischemia; NOS3 gene; Nitric Oxide; Norway; Oxidative Stress; Pathway interactions; Peptide Mapping; Peptides; Personal Satisfaction; Phenotype; Phosphorylation; Phosphorylation Site; Play; Production; Proteins; Proteomics; Rat Strains; Rattus; Recombinants; Research Personnel; Resistance; Role; Signal Transduction; Site; Sodium Chloride; Sprague-Dawley Rats; Superoxides; Technology; Testing; United States; Work; Yang; base; cost; design; domain mapping; human NOS3 protein; indexing; inhibitor/antagonist; insight; normotensive; novel; programs; protein protein interaction; tetrahydrobiopterin

Our understanding of the mechanisms mediating resistance to myocardial ischemia remains unclear. To determine mechanisms for increasing resistance to ischemia, we subjected hearts from Brown Norway (BN/Mcw) and Dahl S (SS/Mcw) rats to global ischemia. Hearts from BN/Mcw rats were more resistance to ischemia than hearts from SS/Mcw rats. Examination of cellular mechanisms revealed that although hearts from both strains exhibited the same levels of endothelial nitric oxide synthase (NOS3) and heat shock protein 90 (hsp90) expression, hearts from BN/Mcw rats generated more nitric oxide (-NO) and less superoxide anion (02'") than hearts from SS/Mcw rats. Basic proteomic studies revealed that the shift in the balance in ¿NOand C>2~ production toward -NO in the BN/Mcw hearts was due, at least in part, to altered heat shock protein 90 (hsp90) association with NOS3 and possibly, with GTP cyclohydrolase I (GTPCH-I). We observed that hsp90 associationwith NOS3 in BN/Mcw hearts was increased nearly 2-fold compared to association in SS/Mcw hearts; and that total biopterin, an analytical index of tetrahydrobiopterin (BH4) was increased in BN/Mcw hearts by 80%. Total biopterin concentrations in hearts from BN/Mcw rats directly correlated with a 70% increase in GTPCH-I protein levels and interestingly, a 2.2-fold increase in association of GTPCH-I with hsp90 compared to the levels in SS/Mcw hearts. These data suggest that hsp90-dependent chaperone activity may play a critical role in mechanisms governing NOS3 function that many consider to be two independent hypotheses: 1) hsp90-dependent signaling modulates NOS3 generation of ¿NO (coupled activity) and OV" (uncoupled activity); and, 2) modulation of BH4levels, which in turn, regulate coupled and uncoupled NOS3 activity. The overall objective of this proposal is to determine the cellular, molecular and enzymatic mechanisms by which hearts from BN/Mcw rats are more resistant to ischemia than hearts from SS/Mcw rats. By elucidating these pathways, findings from this proposal will provide new, fundamental insight into how hsp90 chaperone activity increases NOS3 -NO generation to increase resistance to ischemia. Observations from these studies may actually unify two hypotheses that many consider competing hypotheses in cardiovascular physiology and may also lead to the development of new strategies for treating ischemic heart disease.

我们对介导心肌缺血抵抗的机制的了解尚不清楚。要确定机制以实现 为了增加对缺血的抵抗力，我们用棕色挪威大鼠(BN/Mcw)和达尔·S(SS/Mcw)大鼠的心脏进行全脑缺血。 BN/MCW大鼠心脏对缺血的抵抗力强于SS/MCW大鼠。细胞机制的研究 研究表明，尽管两种菌株的心脏表现出相同的内皮型一氧化氮合酶(NOS3)和热休克水平 蛋白90(HSP90)的表达，BN/MCW大鼠心脏产生更多的一氧化氮(-NO)和更少的超氧阴离子(02‘“) 心脏取自SS/MCW大鼠。基础蛋白质组学研究表明，NO和C&gt；2~产生的平衡向-NO转变 BN/MCW心脏至少部分是由于改变的热休克蛋白90(HSP90)与NOS3相关，可能与GTP相关 环水解酶I(GTPCH-I)。我们观察到，在BN/MCW心脏中，HSP90与NOS3的关联增加了近2倍 与SS/MCW心脏的相关性；以及总生物蝶呤，四氢生物蝶呤(BH4)的分析指数在 BN/MCW心脏增加80%。BN/MCW大鼠心脏总生物蝶呤浓度与GTPCH-I升高70%直接相关 有趣的是，与SS/MCW心脏中的水平相比，GTPCH-I与HSP90的相关性增加了2.2倍。 这些数据表明，依赖于HSP90的伴侣活性可能在调控NOS3功能的机制中发挥关键作用 许多人认为是两个独立的假设：1)依赖HSP90的信号调节NOS3产生NO(偶联 2)调节BH4水平，进而调节偶联和非偶联NOS3的活性。 这项提议的总体目标是确定BN/MCW心脏的细胞、分子和酶机制 与SS/MCW大鼠的心脏相比，大鼠对缺血的抗性更强。通过阐明这些途径，这项提案的发现将 提供有关HSP90伴侣活性如何增加NOS3-NO生成以增强抵抗力的新的、基本的见解 缺血症。这些研究的观察结果实际上可能统一了两个假设，许多人认为这两个假设是相互竞争的 心血管生理学，还可能导致开发治疗缺血性心脏病的新策略。