Plasminogen activator inhibitor type-1 (PAI-1) is the primary inhibitor of tissue-type and urokinase-type plasminogen activators and play integral role in the control of the fibrinolytic system. High plasma PAI-1 level is a risk factor for thrombotic diseases such as myocardial infraction, stroke, and venous thromboembolism. PAI-1 is an early response gene known to be activated by hormones, cytokines, lipoprotein, angiotensin II, phorbol ester, and reactive oxygen species (ROS). The purpose of the present study was to establish whether or not an enhanced oxidative stress involving endothelial injury, activation of coagulation, and inflammatory reaction would induce an expression of PAI-1, t-PA, uPA or uPAR.

Human endothelial cells were treated with hydrogen peroxide (100, 200 µM) , sodium nitroprusside (1mM) and oxidative stress were determined by lucigenin chemiluminescence and 2’7’ dichlorofluorescein (DCF) fluorescence, respectively. PAI-1, tPA, uPA and uPAR transcription were determined by transfection of PAI-1, t-PA, uPA and uPAR promoter linked to a luciferase reporter. Ap1, Sp1 and NFκB activity was determined by an electrophoresis mobility shift assay. Expression of PAI-1, t-PA,uPA and uPAR in vascular endothelial cells studied using microarray assay and confirmed by real time PCR.

This work was supported by the Polish Committee for Scientific Research (KBN) Grant number 2PO5A 141 29 and a Medical University Grant 502-16-300, 502-16-195.

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