MECHANISMS OF ALCOHOL INDUCED ENDOTHELIAL FIBRINOLYSIS

描述：(改编自《调查者摘要》)流行病学 研究表明，适量饮酒(每天1-2杯)可以减少 心血管死亡的风险以及这对心脏保护的好处 可能部分是通过增加纤溶作用来调节的。内皮细胞 (ECS)合成t-PA、u-PA、PAI-1及PA和纤溶酶原受体(Rs) (PMG)(PARS PmgRs)，并维持正常止血和纤溶 通过调节合成和络合物激活EC结合的PMG 这些组件的交互作用。这些EC组件中的更改 /系统性因素的相互作用，即酒精将促进 血栓形成或促进血栓溶解。调查人员已经证明，Low 乙醇(0.1%)可诱导短期(30分钟)和长期持续的双相 (12小时)培养的内皮细胞纤溶功能增强。这些项目的总体目标是 修订后的研究旨在确定/定义分子调控机制。 低乙醇诱导的潜在活性、相互作用和 培养的人EC产生的成分(Pas和Rs)的表达 血管内皮细胞纤溶活性在短期内快速增加，而长期持续增加。 研究将包括短-(无PAS-Rs合成)与 长期(新的PAS/Rs合成)EC结合PMG激活(目标1)；结合 活性(Kd、Bmax)和u-PAR、t-PAR和PmgRs mRNAs的变化(使用 T-PA[Annexin II]和PMG[Annexin II]候选受体的反义探针 和α-烯醇酶]和RPA)，包括电位的测定 乙醇对这些受体类型的转录调控(转录启动 测试)(目标2)；以及最终确定乙醇响应区 T-PA和u-PA基因的启动子和5-侧翼区，包括 它们各自的乙醇诱导转录的初步鉴定 因素(启动子缺失分析、瞬时转染法、EMSA)(目标3)。 从这些研究中收集的结果将提供重要的新见解 使我们更好地理解基因上调背后的机制 低乙醇对EC介导的纤溶活性的影响 基因水平。增加EC-纤溶将大大降低风险 血栓形成、冠心病和相关的动脉粥样硬化血栓事件导致心肌梗死和 将提供一个定义明确的分子基础来部分解释 心脏保护益处和降低心血管死亡风险 与适度饮酒有关。

DESCRIPTION: (Adapted from the Investigator's Abstract) Epidemiologic studies indicate that moderate alcohol consumption (1-2 drinks/day) reduces the risk of cardiovascular mortality and that this cardioprotective benefit may be mediated, in part, by an increase in fibrinolysis. Endothelial cells (ECs) synthesize t-PA, u-PA, PAI-1 and Receptors (Rs) for PA and plasminogen (Pmg) (PARs PmgRs) and maintain normal hemostasis and fibrinolysis by activating EC-bound Pmg through the regulated synthesis and complex interactions of these components. Alterations in these EC components /interactions by systemic factors, i.e. alcohol will promote either thrombosis or facilitate clot lysis. The investigators have shown that low ethanol (<0.1%) induces a biphasic short (<30 min) and long term sustained (12 hr) increase in cultured EC fibrinolysis. The overall goal of these revised studies is ti identify/define the molecular regulatory mechanism underlying low ethanol-induced effects on the activity, interaction, and expression of cultured human EC-produced components (Pas and Rs) resulting in rapid short vs sustained long term increases in EC-fibrinolytic activity. Studies will include kinetic analysis of short-(no Pas Rs synthesis ) vs long term (new Pas/Rs synthesis) EC-bound Pmg activation (Aim 1); binding activity (Kd, Bmax) and changes in u-PAR, t-PAR, and PmgRs mRNAs (using antisens probes for candidate Rs for t-PA [annexinII] and Pmg [annexin II and alpha -enolase] and RPAs), including determination of potential transcriptional regulation of these R types by ethanol (transcription run-on assay) (Aim 2) ; and finally identification of ethanol responsive regions in the promoter and 5-flanking regions of the t-PA and u-PA genes, including initial identication of their respective ethanol-inducible transcription factors (promoter deletion analysis, transient transfection, EMSA) (Aim 3). Results gleaned from these studies will provide significant new insights into our understanding of the mechanisms underlying the upregulation of EC-mediated fibrinolytic activity by low ethanol, at both, the molecular and gene levels. Increased EC-fibrinolysis will substantially decrease the risk for thrombosis, CAD and associated atherothrombotic events leading to MI and will provide a well defined molecular basis to explain, in part, the cardioprotective benefit and reduced risk of cardiovascular mortality associated withe moderate alcohol consumption.