Moderate alcohol or red wine consumption (1-4 drinks/day) reduces the risk for CHD-related mortality. This cardioprotection may be due, in part, to increased fibrinolysis. Endothelial cells (ECs) synthesize t-PA, u-PA, PAI-1 and receptors (Rs) for PAs and plasminogen (Pmg) (PARs, PmgRs) and maintain normal hemostasis/fibrinolysis by activating R-bound Pmg through the regulated synthesis/interactions of these fibrinolytic components. Changes in these EC components/interactions by systemic factors (such alcohol, wine components, in particular polyphenols) that increase fibrinolysis will reduce the risk for thrombosis, atherosclerosis/CHD and the atherothrombotic consequences of MI. We have shown that ethanol/polyphenols increase fibrinolysis in cultured human ECs. The overall goal of these studies is to further identify/define the molecular regulatory mechanisms by which low ethanol/polyphenols (catechin, quercetin) affect the activity/expression of EC PAs, PARs and PmgRs, in vitro and in vivo, resulting in increased EC fibrinolysis. Studies will include effects on: expression of PAs_ARs/PmgRs antigen/mRNA in vivo in mouse aortic endothelium, including direct effects of increased fibrinolysis, in vivo, on clot lysis and inhibition of atherosclerosis in wild type and genetically deficient mice (Aim 1); in vivo and in vitro cross-talk between induced increased fibrinolysis and increased bioavailability of NO, including early activation of cellular kinases (i.e. MAPKs) (Aim 2); changes in expression of EC PARs/PmgRs activity/levels/mRNA (in cultured human coronary artery ECs), including individual R contribution to total ligand binding (using R-specific antisense oligonucleotides), regulation of Rs gene expression (transcriptional and/or post-transcriptional) and other PA- induced effects on PARs/PmgRs expression (in cultured PA-deficient mouse aortic EC) (Aim 3) and; identification of ethanol/polyphenol responsive cis-acting elements in the t-PA and u-PA gene promoters, including their ethanol-/polyphenol-inducible transcription factors (Aim 4). Results gleaned from these studies will provide new insights into the molecular mechanisms by which ethanol/polyphenols regulate EC fibrinolysis and contribute to the cardioprotection attributed to moderate alcohol/red wine consumption. An increased understanding of the mechanisms by which these compounds effectively afford cardioprotection will facilitate future development of new therapeutic approaches/strategies that may be widely applied to reduce the overall population risk for CHD-related mortality.
