Thrombin regulation of mRNA levels of tissue plasminogen activator and plasminogen activator inhibitor-1 in cultured human umbilical vein endothelial cells.
Cultured human umbilical vein endothelial cells release tissue plasminogen activator (t-PA) and type 1 plasminogen activator inhibitor (PAI-1) in response to alpha thrombin stimulation. In order to study the mechanisms of thrombin stimulation, we measured changes in levels of mRNA for t-PA and PAI-1 following exposure of endothelial cells to 3 U/mL alpha thrombin. Alpha thrombin causes a significant and time-dependent increase in the mRNA levels of both t-PA and PAI-1. Catalytically inactivated diisofluorophosphate (DIP) treated thrombin and alpha thrombin pretreated with hirudin do not alter t-PA and PAI-1 mRNA levels. We conclude that the increased secretion of t-PA and PAI-1 by human umbilical vein endothelial cells in response to alpha thrombin is mediated at least partially through an increase in mRNA levels. In addition, an active thrombin catalytic site is required for these increases in mRNA to occur.
Dichek
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Department of Medicine, Massachusetts General Hospital, Boston 02114.
Quertermous
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D001779:Blood Coagulation Factors / Q000235:genetics; D015152:Blotting, Northern; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000502:physiology*; D004731:Endotoxins / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation / Q000187:drug effects; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D066298:In Vitro Techniques; D015849:Plasminogen Inactivators; D012333:RNA, Messenger / Q000235:genetics; D013917:Thrombin / Q000494:pharmacology*; D010959:Tissue Plasminogen Activator / Q000037:antagonists & inhibitors / Q000235:genetics*
D001779:Blood Coagulation Factors; D004731:Endotoxins; D006023:Glycoproteins; D015849:Plasminogen Inactivators; D012333:RNA, Messenger; D013917:Thrombin; D010959:Tissue Plasminogen Activator