MECHANISMS OF ALCOHOL INDUCED ENDOTHELIAL FIBRINOLYSIS

酒精诱导内皮纤维蛋白溶解的机制

• 批准号: 6371447
• 负责人: FRANCOIS M BOOYSE
• 金额: $ 37.1万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-17 至 2003-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6371447
• 关键词: annexins; cardiovascular disorder prevention; chemical kinetics; chemoprevention; disease /disorder proneness /risk; drug interactions; ethanol; fibrinolysis; gene expression; human tissue; plasmin; plasminogen; plasminogen activator; plasminogen activator inhibitors; protein biosynthesis; receptor; tissue /cell culture; vascular endothelium

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.

描述：（改编自研究者摘要）流行病学 研究表明，适量饮酒（1-2杯/天） 心血管死亡的风险和这种心脏保护益处 可能部分由纤维蛋白溶解的增加介导。 内皮细胞 (ECs)合成t-PA、u-PA、派-1以及PA和纤溶酶原受体（Rs (Pmg)（PARs Pmgr）并通过以下方式维持正常止血和纤溶： 通过调节合成和复合物激活EC结合的Pmg 这些组件的相互作用。 这些EC组件的变更 /系统因素的相互作用，即酒精会促进 血栓形成或促进凝块溶解。 调查显示，低 乙醇（<0.1%）诱导双相短期（<30分钟）和长期持续 (12 hr）培养的EC纤维蛋白溶解增加。 这些活动的总体目标 修订的研究是为了确定/定义分子调节机制 潜在的低乙醇诱导的活性，相互作用， 表达培养的人EC产生的组分（Pas和Rs）， EC纤溶活性的快速短期增加与持续长期增加。 研究将包括短链（无Pas Rs合成）与短链（无Pas Rs合成）的动力学分析。 长期（新的Pas/Rs合成）EC结合的Pmg激活（目的1）;结合 活性（Kd，Bmax）和u-PAR，t-PAR和PmGRs mRNA的变化（使用 t-PA [膜联蛋白II]和Pmg [膜联蛋白II]候选Rs的反义探针 和α-烯醇化酶]和RPAs），包括测定 乙醇对这些R型的转录调节（转录连续 试验）（目的2）;最后鉴定乙醇反应区， t-PA和u-PA基因的启动子和5-侧翼区，包括 它们各自乙醇诱导转录的初步鉴定 因子（启动子缺失分析、瞬时转染、EMSA）（目的3）。 从这些研究中收集的结果将提供重要的新见解 使我们了解到， 低浓度乙醇对内皮细胞介导的纤溶活性的影响 基因水平。 增加EC-纤维蛋白溶解将大大降低风险 血栓形成、CAD和导致MI的相关动脉粥样硬化血栓形成事件， 将提供一个明确的分子基础来解释，在某种程度上， 心脏保护获益和降低心血管死亡风险 与适度饮酒有关。