To transdifferentiate human hypertrophic scar fibroblasts induced by connective tissue growth factor mediated transforming growth factor-β1, in vitro
Author(s): LI Zhe, LI Shi-rong, LIU Jian-yi, DAI Xia, TAO Ling
Pages: 49-52
Year: 2009 Issue: 1
Journal: Chinese Journal of Burns
Keyword: Transforming growth factor beta 1; Fibroblasts; Transdifferentiation; Connective tissue growth factor; Hypertrophic sear;
Abstract: Objective To investigate the role of connective tissue growth factor(CTGF) induced TGF-β1 in the transdifferentiation of human hypertrophic scar fibroblast(HSFb). Methods Human hy-pertrophic scar fibroblasts were cultured in vitro, 5 cell samples were stimulated with TGF-β(0, 2.5, 5.0, 7.5, 10.0 ng/mL, respectively)for 48 hours; other cell samples were divided into: normal control (NC) group, CTGF group(with addition of 10 ng/mL rhCTGF into culture medium), TGF-β1 group(with addition of 10 ng/mL TGF-β1 into culture medium), CTGF ASODN group(with addition of 10% FBS-DMEM after transfection of CTGF ASODN), CTGF ASODN + TGF-β1 group(with addition of 10 ng/mL TGF-β1 after transfection of CTGF ASODN). Expression of CTGF was determined by Western blotting with stimulation of different concentration of TGF-β1. Expression of a-smooth muscle actin(α-SMA) was measured by Western blotting. Positive cell rate of α-SMA was examined by flow cytometry. Results With stimulation of 10.0 ng/mL TGF-β1, the expression of CTGF was obviously higher than that of non-stimulation (P <0.05). Ex-pression of α-SMA in the CTGF group and the TGF-β1 group was obviously higher than that in NC group (P<0.01),while there was no obvious difference among NC, CTGF ASODN, CTGF ASODN + TGF-β1 groups(P >0.05). The positive cell rate of α-SMA in NC, CTGF, TGF-β1, CTGF ASODN, CTGF ASODN +TGF-β1 groups was(10. 8±2.8)%, (29.1±4.0)%, (28.7±4.8)%, (10.7±2.3)%, (14.3±2.9) %, respectively, which was similar to expression of α-SMA on statistic analysis. Conclu-sions CTGF is one of the most important downstream effortors for TGF-β1 in inducing the transdifferentia-tion of HSFb.
Pages: 49-52
Year: 2009 Issue: 1
Journal: Chinese Journal of Burns
Keyword: Transforming growth factor beta 1; Fibroblasts; Transdifferentiation; Connective tissue growth factor; Hypertrophic sear;
Abstract: Objective To investigate the role of connective tissue growth factor(CTGF) induced TGF-β1 in the transdifferentiation of human hypertrophic scar fibroblast(HSFb). Methods Human hy-pertrophic scar fibroblasts were cultured in vitro, 5 cell samples were stimulated with TGF-β(0, 2.5, 5.0, 7.5, 10.0 ng/mL, respectively)for 48 hours; other cell samples were divided into: normal control (NC) group, CTGF group(with addition of 10 ng/mL rhCTGF into culture medium), TGF-β1 group(with addition of 10 ng/mL TGF-β1 into culture medium), CTGF ASODN group(with addition of 10% FBS-DMEM after transfection of CTGF ASODN), CTGF ASODN + TGF-β1 group(with addition of 10 ng/mL TGF-β1 after transfection of CTGF ASODN). Expression of CTGF was determined by Western blotting with stimulation of different concentration of TGF-β1. Expression of a-smooth muscle actin(α-SMA) was measured by Western blotting. Positive cell rate of α-SMA was examined by flow cytometry. Results With stimulation of 10.0 ng/mL TGF-β1, the expression of CTGF was obviously higher than that of non-stimulation (P <0.05). Ex-pression of α-SMA in the CTGF group and the TGF-β1 group was obviously higher than that in NC group (P<0.01),while there was no obvious difference among NC, CTGF ASODN, CTGF ASODN + TGF-β1 groups(P >0.05). The positive cell rate of α-SMA in NC, CTGF, TGF-β1, CTGF ASODN, CTGF ASODN +TGF-β1 groups was(10. 8±2.8)%, (29.1±4.0)%, (28.7±4.8)%, (10.7±2.3)%, (14.3±2.9) %, respectively, which was similar to expression of α-SMA on statistic analysis. Conclu-sions CTGF is one of the most important downstream effortors for TGF-β1 in inducing the transdifferentia-tion of HSFb.
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