PROTECTION FROM CARDIAC REPERFUSION INJURY BY ETHANOL

申请者摘要：适度使用乙醇与保护相关 对抗致命的冠状动脉事件。然而，除了下降之外， 冠状动脉事件的发生率，事件后恢复的改善可能是 乙醇对预防冠状动脉致死结局的重要作用 疾病。减轻缺血后再灌注损伤的机制之一是通过 这种心肌恢复，从而存活率可能会改善后， 冠脉事件。初步研究表明，适量使用乙醇会减弱 激活腺苷受体引起的豚鼠再灌注损伤 红心。假设I和II将针对问题1-5进行测试： I.适度使用乙醇对缺血后心肌的保护作用 再灌流 受伤。 1.适量的酒精摄入是否能改善功能和代谢 回收 在再灌流期间？ 2.在缺血再灌注后，适量使用乙醇 减少心脏灌流后心肌细胞坏死量 活体心肌梗死面积？ 3.适量使用乙醇是否能防止钙超载 缺血后再灌流？ 二、适量乙醇对再灌流的保护作用 伤害 是腺苷受体介导的蛋白激酶C的结果 易位。 4.乙醇的保护作用是否被腺苷A1、A2、 和/或A3受体拮抗剂？ 5.乙醇引起的心肌细胞内Delta和epsilon PKC易位吗？ 暴露的心脏和对照组？ 离体豚鼠心脏(10%乙醇喂养6周)将接受 缺血再灌流。能源消耗和回收将通过以下指标进行评估 31P-MRS；肌酸激酶释放引起的心肌细胞坏死；胞浆内钙离子的释放 Indo-1荧光；免疫荧光法测定PKC同工酶易位 定位和蛋白质印迹分析。在活体内，梗死面积将是 左冠状动脉闭塞-再灌流后测量。 本研究的最终目标是：1)建立机制 适量饮酒与心脏病之间的正相关 保健和2)协助制定治疗干预措施， 在有致命冠状动脉事件风险的患者中模拟这种效果，这些患者需要 选择性或紧急再灌流。

APPLICANT'S ABSTRACT: Moderate ethanol use is associated with protection against fatal coronary events. However, in addition to a decreased incidence of coronary events, improved recovery following an event may be important in ethanol's prevention of fatal outcomes due to coronary artery disease. Attenuation of post-ischemic reperfusion injury is a mechanism by which myocardial recovery, and thereby survival, may be improved following a coronary event. Pilot studies suggest that moderate ethanol use attenuates reperfusion injury through activation of adenosine receptors in guinea pig hearts. Hypotheses I and II will be tested addressing questions #1-5: I. Moderate ethanol use protects against myocardial post-ischemic reperfusion injury. 1. Does moderate ethanol use improve functional and metabolic recovery during reperfusion? 2. Following post-ischemic reperfusion, does moderate ethanol use decrease the amount of myocyte necrosis in perfused hearts and infarct size in vivo? 3. Does moderate ethanol use protect against Ca2+ overload during post-ischemic reperfusion? II. The protective effect of moderate ethanol use against reperfusion injury is the result of adenosine receptor mediated protein kinase C translocation. 4. Is ethanol's protective effect abolished by adenosine A1, A2, and/or A3 receptor antagonists? 5. Are delta and epsilon PKC translocated in myocytes from ethanol exposed hearts versus controls? Isolated guinea pig hearts (fed 10% ethanol for 6 weeks) will undergo ischemia - reperfusion. Energy depletion and recovery will be assessed with 31P-MRS; myocyte necrosis by creatine kinase release; cytosolic CA2+ by indo-1 fluorescence; and PKC isozyme translocation by immunofluorescence localization and western blot analysis. In vivo, infarct size will be measured after occlusion-reperfusion of the left coronary artery. The ultimate goals of this research are 1) to establish the mechanisms underlying the positive association between moderate alcohol use and cardiac health and 2) aid in the development of therapeutic interventions which mimic this effect in patients at risk for fatal coronary events who require elective or emergent reperfusion.