G Proteins and Phospholipase C Mediate Thrombin-lnduced Generation of Plasminogen Activator Inhibitor-1 from Vascular Smooth Muscle Cells

Abstract

The present study investigated transcellular signalling mechanism involved in thrombin-induced production of plasminogen activator inhibitor-1 (PAI-1) in cultured vascular baboon aortic smooth muscle cells (BASMC). Treatments with thrombin dose-dependently increased the steady state levels of PAI-1 mRNA and the generation of PAI-1 antigen from BASMC. Thrombin receptor-activating peptide mimicked the effect of thrombin on the generation of PAI-1. Sodium fluoride (1 m M) stimulated PAI-1 generation from BASMC. Pertussis toxin dose-dependently suppressed thrombin-induced increase of PAI-1 generation. Treatment with 5 m M neomycin, 10 µ M U73122 or 1 µ M calphostin C blocked thrombin-induced PAI-1 generation. Phorbol myristate acetate at 10 n M for 3 h strongly stimulated the generation of PAI-1 from BASMC. Forskolin (100 µ M)or 8-bromo-cAMP (100 µ M)suppressed thrombin-induced PAI-1 generation. The responses of quiescent BASMC to thrombin or the inhibitors on PAI-1 generation were comparable to that of growing cells. The results of the present study suggest that pertussis toxin-sensitive G proteins and a phospholipase C are involved in thrombin-induced generation of PAI-1 in BASMC, which may transmit signals from occupied thrombin receptor to protein kinase C and thereby increase the generation of PAI-1. Elevated levels of intracellular cAMP may negatively regulate the generation of PAI-1 from vascular SMC.