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Engineered tissue based detection of PAR1 inhibition in thrombin‐induced profibrotic phenotypes of cardiac fibroblasts
Author(s) -
Wakatsuki Tetsuro,
Peterson Matthew,
Strande Jennifer L
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.1086.4
Subject(s) - thrombin , procollagen peptidase , cardiac fibrosis , fibroblast , contractility , endocrinology , chemistry , fibrosis , medicine , ctgf , zymography , receptor , microbiology and biotechnology , biology , growth factor , matrix metalloproteinase , biochemistry , in vitro , platelet
Study objective To determine the effects of PAR1 (Protease‐activated receptor) activation on profibrotic response in cardiac fibroblast (CF) using isolated cells and cardiac connective tissue equivalents (CCTEs). Methods Rat CFs were mixed with collagen to generate CCTEs, and they were treated for 24 hrs with thrombin and/or a PAR1 antagonist. Their tissue stiffness and cellular contractility were measured using a high‐throughput mechanical measurement system. Treating with thrombin and/or PAR1 antagonists, the protein expression/activity of procollagen (type I), TGFb, MMP2/9, ERK1/2 were analyzed using ELISA, immunoblotting and zymography. Results Thrombin treatment stiffened and contracted CCTE by 38% and 25% respectively over controls (n=15). In isolated CFs, thrombin activated ERK1/2 as well as increased MMP2/9 activities and procollagen and TGFb secretion over the control. PAR1 and ERK1/2 inhibitors attenuated these effects. Conclusions Thrombin stimulated CCTEs to increase tissue stiffness and cell contractility. This profibrotic CCTE remodeling was correlated with ERK1/2 activation, increased TGFb and procollagen secretion and activation of MMP2/9. The anti‐fibrotic effects of PAR1 inhibition strongly suggests that PAR1 signaling plays a critical role in ERK1/2 mediated thrombin‐induced cardiac fibrosis.