Protection From Cardiac Reperfusion Injury by Ethanol

描述(由申请人提供)：适度饮酒已被证明为 在众多的流行病学研究中，减少冠心病。我们建议 适量的酒精摄入通过增加 心肌细胞中epsilonPKC蛋白的表达。为了检验这一假设，我们 将使用多种方法来检测缺血-再灌注的阻力 在饮用水中饮用乙醇至少12周会对心脏造成伤害。在……里面 此外，我们将研究epsilonPKC功能在 乙醇介导的心脏保护使用我们的常规技术 具有以下特定目标的实验室：目标A：检测epsilonPKC酶 乙醇灌胃小鼠心脏的活性和亚细胞定位 年龄匹配的对照组。我们计划确定乙醇引起的epsilonPKC的变化 成人肌动蛋白的功能及其在亚细胞间的分布 乙醇灌胃小鼠的心肌细胞和左心室组织中 年龄匹配的对照组使用免疫荧光染色，共聚焦显微镜， 免疫沉淀和免疫印迹技术。 目的B：确定急性同工酶选择性抑制epsilonPKC 功能阻断持续的乙醇介导的心脏保护。我们将研究 多肽调节剂对蛋白激酶C同工酶转位和功能的影响 将其急性导入培养的成人心肌细胞或完整心脏 慢性乙醇诱导的脑缺血再灌注耐受与蛋白激酶C 与其他信号蛋白的相互作用。目标C：调查 适量饮酒对心脏功能和抗药性的影响 EpsilonPKC基因敲除小鼠的缺血再灌注损伤。我们将使用成人 确定心肌细胞和完整心脏是否具有心脏保护作用 在epsilonPKC基因敲除小鼠中对酒精喂养的反应以及是否 乙醇介导的相关信号通路的调节被 不存在epsilon PKC。这项研究的一个总体目标是理解 慢性中度酒精所致心脏保护的细胞机制 消费。第二个目标是确定持续的治疗靶点 对冠心病的保护，不需要摄入酒精 由于对酒精滥用和潜在不良影响的担忧 人类的其他器官系统。

DESCRIPTION (provided by applicant): Moderate alcohol intake has been shown to reduce coronary heart disease in numerous epidemiological studies. We propose that moderate ethanol consumption causes cardioprotection by increasing epsilonPKC protein expression in cardiac myocytes. To test this hypothesis, we will use multiple approaches to examine resistance of ischemia-reperfusion injury in hearts receiving ethanol in drinking water for at least 12 weeks. In addition, we will investigate the requirement for epsilonPKC function in ethanol-mediated cardioprotection using techniques routinely available in our laboratory with the following specific aims: Aim A: Examine epsilonPKC enzyme activity and subcellular localization in hearts from ethanol-fed mice and age-matched controls. We plan to identify ethanol-induced changes in epsilonPKC kinase function and distribution among subcellular compartments in adult cardiac myocytes and in left ventricular tissue from ethanol-fed mice and age-matched controls using immunofluorescence staining, confocal microscopy, immunoprecipitation, and western blotting techniques. Aim B: Determine whether acute isozyme-selective inhibition of epsilonPKC function blocks sustained ethanol-mediated cardioprotection. We will examine the effects of peptide modulators of PKC isozyme translocation and function introduced acutely into cultured adult cardiac myocytes or intact hearts on chronic ethanol-induced resistance to ischemia-reperfusion injury and PKC interactions with other signaling proteins. Aim C: Investigate the effects of moderate alcohol consumption on cardiac function and resistance to ischemia-reperfusion injury in epsilonPKC knockout mice. We will use adult cardiac myocytes and intact hearts to determine whether cardioprotection develops in epsilonPKC knockout mice in response to ethanol feeding and whether ethanol-mediated regulation of related signaling pathways is altered by the absence of epsilonPKC. One overall goal of this research is to understand the cellular mechanisms of cardioprotection mediated by chronic moderate alcohol consumption. A second goal is to identify therapeutic targets for sustained protection against coronary heart disease that do not require ethanol ingestion because of concerns regarding alcohol abuse and potential adverse effects on other organ systems in humans.