redox-induced src kinase and caveolin-1 signaling in tgf-β1-initiated smad23 activation and pai-1 expressionredox-induced src激酶和caveolin-1信号tgf-β1-initiated smad23激活和pai-1表达式.pdfVIP

Redox-Induced Src Kinase and Caveolin-1 Signaling in TGF-b1-Initiated SMAD2/3 Activation and PAI-1 Expression 1 1 1 1 2 Rohan Samarakoon , Subhanir S. Chitnis , Stephen P. Higgins , Craig E. Higgins , Joan C. Krepinsky , Paul J. Higgins1* 1 Center for Cell Biology and Cancer Research, Albany Medical College, Albany, New York, United States of America, 2 Division of Nephrology, McMaster University, Hamilton, Ontario, Canada Abstract Background: Plasminogen activator inhibitor-1 (PAI-1), a major regulator of the plasmin-based pericellular proteolytic cascade, is significantly increased in human arterial plaques contributing to vessel fibrosis, arteriosclerosis and thrombosis, particularly in the context of elevated tissue TGF-b1. Identification of molecular events underlying to PAI-1 induction in response to TGF-b1 may yield novel targets for the therapy of cardiovascular disease. Principal Findings: Reactive oxygen species are generated within 5 minutes after addition of TGF-b1 to quiescent vascular smooth muscle cells (VSMCs) resulting in pp60c-src activation and PAI-1 expression. TGF-b1-stimulated Src kinase signaling sustained the duration (but not the initiation) of SMAD3 phosphorylation in VSMC by reducing the levels of PPM1A, a recently identified C-terminal SMAD2/3 phosphatase, thereby maintaining SMAD2/3 in an active state with retention of PAI- 1 transcription. The markedly increased PPM1A levels in triple Src kinase (c-Src, Yes, Fyn)-null fibroblasts are consistent with reductions in both SMAD3 phosphorylation and PAI-1 expression in response to TGF-b1 compared to wild-type cells. Activation of the Rho-ROCK pathway was mediated by Src kinases