Hsp90, NOS3 and Cardioprotection

Hsp90、NOS3 和心脏保护

• 批准号: 7879747
• 负责人: Yang Shi
• 金额: $ 21.72万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7879747
• 关键词: Anabolism; Biopterin; Cardiovascular Physiology; Cessation of life; 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; 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.

我们对介导对心肌缺血抗性的机制的理解尚不清楚。确定机制 越来越多的缺血性抗性，我们将心脏从北挪威（BN/MCW）和Dahl S（SS/MCW）大鼠进行到全球性缺血。 来自BN/MCW大鼠的心脏比SS/MCW大鼠的心脏更具缺血性。 cell 揭示了尽管两种菌株的心脏都暴露了相同水平的内皮一氧化氮合酶（NOS3）和热休克 蛋白质90（HSP90）表达，BN/MCW大鼠的心脏比一氧化氮（-NO）和超氧化物阴离子（02'”）比 SS/MCW老鼠的心。基本的蛋白质组学研究表明，noand c> 2〜的平衡的变化向-no中 BN/MCW心脏至少部分是由于与NOS3的相关性，并且可能与NOS3的关联，并且与GTP发生了变化 环氢酶I（GTPCH-I）。我们观察到BN/MCW心脏中的HSP90与NOS3的关联增长了近2倍 与SS/MCW心脏中的关联相比；并且总生物蛋白是四氢无菌蛋白酶（BH4）的分析指数 BN/MCW心增加80％。 BN/MCW大鼠心脏中生物蛋白的总浓度直接与GTPCH-I的增长直接相关 蛋白质水平，有趣的是，与SS/MCW心脏的水平相比，GTPCH-I与HSP90的缔合增加了2.2倍。 这些数据表明，HSP90依赖性伴侣活动可能在管理NOS3功能的机制中起关键作用， 许多人认为是两个独立的假设：1）HSP90依赖性信号传导调节NOS3的生成„ no（耦合） 活性）和ov“（未偶联活性）; 2）BH4Levels的调节，进而调节耦合和未耦合的NOS3活性。 该提案的总体目的是确定BN/MCW的心脏的细胞，分子和酶促机制 大鼠比SS/MCW大鼠的心脏更耐缺血。通过阐明这些途径，该提议的发现将 提供有关HSP90伴侣活动如何增加NOS3的新的基本见解 缺血。这些研究的观察实际上可能统一了两个假设，许多假设考虑了竞争的假设 心血管生理学，也可能导致制定治疗缺血性心脏病的新策略。

项目成果

Mesenteric nitric oxide and superoxide production in experimental necrotizing enterocolitis.

实验性坏死性小肠结肠炎中肠系膜一氧化氮和超氧化物的产生。

• DOI: 10.1016/j.jss.2009.07.028
• 发表时间: 2010
• 期刊: The Journal of surgical research
• 作者: Whitehouse,JillS;Xu,Hao;Shi,Yang;Noll,LeAnne;Kaul,Sushma;Jones,DeronW;PritchardJr,KirkwoodA;Oldham,KeithT;Gourlay,DavidM
• 通讯作者: Gourlay,DavidM

Increased resistance to LPS-induced myocardial dysfunction in the Brown Norway rats versus Dahl S rats: roles of inflammatory cytokines and nuclear factor kappaB pathway.

• DOI: 10.1097/shk.0b013e3181b7819e
• 发表时间: 2010-03
• 期刊: Shock (Augusta, Ga.)
• 作者: Du J;An J;Wei N;Guan T;Pritchard KA Jr;Shi Y
• 通讯作者: Shi Y

Differential sensitivity to LPS-induced myocardial dysfunction in the isolated brown Norway and Dahl S rat hearts: roles of mitochondrial function, NF-κB activation, and TNF-α production.

• DOI: 10.1097/shk.0b013e31823f146f
• 发表时间: 2012-03
• 期刊: Shock (Augusta, Ga.)
• 作者: An J;Du J;Wei N;Guan T;Camara AK;Shi Y
• 通讯作者: Shi Y