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Ethanol Transcriptionally Upregulates t‐PA and u‐PA Gene Expression in Cultured Human Endothelial Cells
Author(s) -
Grenett Hernan E.,
Aikens Michael L.,
Torres Jose A.,
Demissie Sossiena,
Tabengwa Edlue M.,
Davis Glenda C.,
Booyse Francois M.
Publication year - 1998
Publication title -
alcoholism: clinical and experimental research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.267
H-Index - 153
eISSN - 1530-0277
pISSN - 0145-6008
DOI - 10.1111/j.1530-0277.1998.tb03878.x
Subject(s) - fibrinolysis , plasminogen activator , umbilical vein , microbiology and biotechnology , luciferase , tissue plasminogen activator , downregulation and upregulation , gene expression , biology , chemistry , transfection , gene , medicine , in vitro , endocrinology , biochemistry
Epidemiological studies have suggested that moderate alcohol consumption reduces the risk of cardiovascular mortality. This cardio‐protective benefit may be mediated, in part, by promoting fibrinolysis through changes in fibrinolytic components and/or activity, resulting in the decreased risk for thrombosis, coronary artery disease, and eventual myocardial infarction. Endothelial cells (ECs) play a pivotal role in maintaining normal hemostasis by regulating fibrinolysis through the synthesis of plasminogen activators (PAS), tissue‐type plasminogen activator (t‐PA), and urokinase‐type plasminogen activator (u‐PA). The studies described herein were conducted to determine whether a single brief preincubation (1 hr, 37°C) of cultured human umbilical vein ECs (HUVECs) with low ethanol (0.1%, v/v), will upregulate t‐PA and/or u‐PA gene expression at the transcriptional level, using a combination of nuclear transcription run‐on assays and transient transfections of cultured HUVECs with the pPA/luc promoter constructs. Nuclear run‐on assays showed ∼2‐ to 3‐fold and ∼6‐ to 7‐fold increase in the transcription of new t‐PA and u‐PA mRNAs, respectively. In addition, transient transfections of cultured HUVECs with the pt‐PA363/luc and pu‐PA236/luc promoter constructs, using lipofectamine, demonstrated ∼4‐ to 6‐fold and ∼6‐ to 9‐fold increase in luciferase activity for t‐PA and U‐PA, respectively. These combined results demonstrate that low ethanol transcription‐ally upregulates both t‐PA and U‐PA gene expression in cultured HUVECs and provides a molecular basis for the ethanol‐induced increase in EC‐mediated fibrinolytic activity that may underlie and contribute, in part, to the cardioprotective benefit associated with moderate alcohol consumption.