title: Thrombin stimulates smooth muscle cell differentiation from peripheral blood mononuclear cells via protease-activated receptor-1, RhoA, and myocardin.
abstract: BACKGROUND
  Smooth muscle precursor cells have previously been reported to reside in bone marrow and in the circulation, but little is currently known regarding the proximate stimuli for smooth muscle cell differentiation of these putative progenitors.
  
  
  OBJECTIVE
  Because local thrombin generation occurs as an initial response to vascular injury, we hypothesized that thrombin may influence the differentiation of circulating smooth muscle progenitor cells.
  
  
  RESULTS
  Peripheral blood mononuclear cells were cultured on type I collagen using a protocol optimized to stimulate smooth muscle cell outgrowth. Thrombin-stimulated upregulation of the transcription factor myocardin and smooth muscle myosin heavy chain, and both were inhibited by hirudin or the RhoA inhibitor Y27632. After 10 days of culture, smooth muscle outgrowth colonies formed, which stained positive for alpha-smooth muscle actin, smooth muscle myosin heavy chain, and calponin, in addition to having a contractile response to 100 nmol/L angiotensin II. Coincubation of peripheral blood mononuclear cells with thrombin, 10 micromol/L protease-activated receptor-1, but not protease-activated receptor-4 activating peptide significantly increased the number of smooth muscle outgrowth colonies formed. Thrombin-induced enhancement of smooth muscle outgrowth colony formation was inhibited by hirudin, Y27632, and an antibody against protease-activated receptor-1.
  
  
  CONCLUSIONS
  These data illustrate a novel thrombin-induced pathway for smooth muscle differentiation from putative smooth muscle progenitors in peripheral blood.
authors: Centre for Research in Vascular Biology, Biosciences Institute, University College Cork, Ireland.
authors: Kenneth
authors: K
authors: Martin
authors: Sharon
authors: S
authors: Weiss
authors: Pat
authors: P
authors: Metharom
authors: Jeffrey
authors: J
authors: Schmeckpeper
authors: Brian
authors: B
authors: Hynes
authors: John
authors: J
authors: O'Sullivan
authors: Noel
authors: N
authors: Caplice
mesh_terms: D000577:Amides / Q000494:pharmacology; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D006629:Hirudins / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D044463:Receptor, PAR-1 / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D013552:Swine; D013917:Thrombin / Q000378:metabolism / Q000494:pharmacology*; D015534:Trans-Activators / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000099251:Myocardin
chemical_list: D000577:Amides; D006629:Hirudins; D018995:Myosin Heavy Chains; D009687:Nuclear Proteins; D011480:Protease Inhibitors; D011725:Pyridines; D044463:Receptor, PAR-1; D013917:Thrombin; D015534:Trans-Activators; D020742:rhoA GTP-Binding Protein; D000099251:Myocardin; C108830:Y 27632