title: [Transforming growth factor-β1 induces differentiation of bone marrow-derived mesenchymal stem cells into myofibroblasts via production of reactive oxygen species].
abstract: OBJECTIVE
  The aim of this study was to investigate the mechanism underlying transforming growth factor-β1 (TGF-β1) induced differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) into myofibroblasts.
  
  
  METHODS
  Primary mouse BMSCs were isolated from bone marrow by flushing the tibias and femurs of mice, and passage 3 to passage 5 of BMSCs were used in the experiments. BMSCs differentiation into myofibroblast was induced by different doses of TGF-β1. In addition, reactive oxygen species (ROS) inhibitor (N-acetylcysteine, NAC) was added to test its effect on the action of TGF-β1. Expressions of BMSCs differentiation parameters, α-smooth muscle actin (α-SMA), collagen α1(I) [Col α1(I)] and collagen α1(III) [Col α1(III)] were measured by real-time quantitative PCR (RT-qPCR) and Western blot analysis. BMSCs were preloaded for 15 min with 2', 7'-dichlorohydrofluorescein diacetate (DCFH-DA), then stimulated with TGF-β1 for different times, and fluorescence of ROS was measured using high content analysis.
  
  
  RESULTS
  TGF-β1 stimulated differentiation of BMSCs into myofibroblasts and up-regulated expression of α-SMA, Col α1(I) and Col α1(III) in a dose-dependent manner, which blocked by ROS inhibitor NAC. In addition, TGF-β1 could induce a significant rapid and transient increase in ROS production in BMSCs, and the effect of TGF-β1 on ROS production was peaked at 30 min.
  
  
  CONCLUSIONS
  TGF-β1 induced differentiation of BMSCs into myofibroblasts via production of ROS.
authors.affiliation: Department of Cell Biology, Municipal Laboratory for Liver Protection and Regulation of Regeneration, Capital Medical University, Beijing 100069, China.
authors.forename: Shuang-shuang
authors.initials: SS
authors.lastname: Jia
authors.affiliation: Department of Cell Biology, Municipal Laboratory for Liver Protection and Regulation of Regeneration, Capital Medical University, Beijing 100069, China.
authors.forename: Wei-yang
authors.initials: WY
authors.lastname: Li
authors.affiliation: Department of Cell Biology, Municipal Laboratory for Liver Protection and Regulation of Regeneration, Capital Medical University, Beijing 100069, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Cell Biology, Municipal Laboratory for Liver Protection and Regulation of Regeneration, Capital Medical University, Beijing 100069, China.
authors.forename: Li-ying
authors.initials: LY
authors.lastname: Li
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation; D003094:Collagen / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D058628:Myofibroblasts / Q000166:cytology*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*
chemical_list: D000199:Actins; D017382:Reactive Oxygen Species; D053773:Transforming Growth Factor beta1; C544013:alpha-smooth muscle actin, mouse; D003094:Collagen