Anesthetic-Induced Cardiac Preconditioning

描述（由申请人提供）：与缺血预处理的强大内源性心脏保护机制相似，麻醉诱导预处理（APC）已成为一种同样有效的心脏保护干预措施，对患者具有更好的风险受益比。在本方案项目周期内，我们确定了APC所涉及的关键要素和机制。心脏保护的核心是我们已经获得的关于挥发性麻醉剂调节线粒体功能的知识。鉴于线粒体在心肌能量学和氧化应激中的基本作用，我们认为它们是APC等保护策略的有希望的靶点。相反，对APC有抗性的疾病状态（例如糖尿病）包含线粒体功能的基本紊乱。 本计划的中心主题是阐明APC的分子机制。具体而言，我们假设，缺血和再灌注后的渗透性转换（PT）孔开放衰减是暴露于挥发性麻醉剂后观察到的许多表型差异的核心。该计划将由两个核心支持的三个相互关联的研究项目组成。项目I（Warltier）将专注于确定与挥发性麻醉剂通过HIF 1a-VEGF-NO <$轴调节NO <$生产相关的关键心脏保护蛋白的激活时间顺序。将确定这些元件的破坏及其对sarcKATp通道活性、神经源性ROS形成以及组织和细胞损伤的影响。项目II（Bosnjak）将阐明挥发性麻醉依赖性PT孔开放调节（APC的关键末端效应器）的机制。它将解决几个因素是至关重要的PT孔在APC中的作用，如线粒体生物能量学和它的蛋白质组和sarcKATp通道的贡献。线粒体生物能量学的计算模型将用于检验与挥发性麻醉剂的作用相关的特定假设。项目III（Kersten）将研究糖尿病动物中APC衰减的相关机制。它将开发一种新的2型糖尿病大鼠模型，在该模型中，我们能够选择性地切换线粒体基因组，以进一步剖析线粒体和eNOS敏感通路在APC受损期间的作用。所有三个项目将得到生化和分子生物学核心（哈德）和蛋白质组学核心（奥利弗）的支持。这些核心将提供最先进的技术，在基因沉默，真实的时间PCR，线粒体蛋白质组，细胞培养，线粒体功能测定，共聚焦显微镜和病理学。 该计划项目代表了一项全面的努力，利用我们在生理学，生物物理学，基因组学，蛋白质组学和计算生物学方面的现有基础设施和计划经验，以促进我们对麻醉剂在器官保护中的细胞和亚细胞作用的理解。我们的研究结果可能对挥发性麻醉药的临床应用产生重大影响。

DESCRIPTION (provided by applicant): Similar to the powerful endogenous cardioprotective mechanism of ischemic preconditioning, anesthetic-induced preconditioning (APC) has emerged as an equally effective cardioprotective intervention with better risk-to-benefit ratio for the patient. During the current cycle of the Program Project we have identified key elements and mechanisms involved in APC. Central to cardioprotection is the knowledge we have gained regarding the regulation of mitochondrial function by volatile anesthetics. Given the fundamental role of mitochondria in myocardial energetics and oxidative stress, we believe that they are a promising target for protective strategies such as APC. In contrast, disease states resistant to APC (e.g. diabetes) contain fundamental disturbances of mitochondrial function. The central theme of this Program Project is to elucidate the molecular mechanisms underlying APC. Specifically, we hypothesize that attenuation of permeability transition (PT) pore opening after ischemia and reperfusion is central to many of the phenotypic differences observed after exposure to volatile anesthetics. This Program will consist of three interrelated research projects supported by two Cores. Project I (Warltier) will focus on defining the temporal sequence of activation of key cardioprotective proteins related to the regulation of NO¿ production via HIF1a-VEGF-NO¿ axis by volatile anesthetics. Disruption of these elements and their effect on sarcKATp channel activity, mitochondrial-derived ROS formation, and tissue and cell injury will be determined. Project II (Bosnjak) will elucidate mechanisms involved in volatile anesthetic-dependent modulation of PT pore opening, a critical end effector of APC. It will address several factors that are critical to the role of the PT pore in APC such as mitochondrial bioenergetics and its proteome and the contribution of sarcKATp channels. Computational models of mitochondrial bioenergetics will be used to test specific hypotheses related to the effects of volatile anesthetics. Project III (Kersten) will investigate mechanisms involved in the attenuation of APC in diabetic animals. It will exploit a novel rat model of type 2 diabetes in which we were able to selectively switch the mitochondrial genome to further dissect the role of mitochondria and eNOS-sensitive pathway during impaired APC. All three Projects will be supported by a Biochemical and Molecular Biology Core (Harder) and a Proteomics Core (Olivier). These Cores will provide state-ofthe- art techniques in gene silencing, real time PCR, mitochondrial proteome, cell cultures, mitochondrial function assays, confocal microscopy and pathology. This Program Project represents a comprehensive effort to leverage our existing infrastructure and programmatic experience in physiology, biophysics, genomics, proteomics, and computational biology to advance our understanding of the cellular and subcellular effects of anesthetics in organ protection. Our findings are likely to have a significant impact in the clinical use of volatile anesthetics.