Zhonghua shao shang za zhi = Zhonghua shaoshang zazhi = Chinese journal of burns · 2009
[To transdifferentiate human hypertrophic scar fibroblasts induced by connective tissue growth factor mediated transforming growth factor-beta 1 in vitro].
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Li Zhe, Li Shi-rong, Liu Jian-yi, Dai Xia, Tao Ling
Аннотация
OBJECTIVE: To investigate the role of connective tissue growth factor (CTGF) induced TGF-beta 1 in the transdifferentiation of human hypertrophic scar fibroblast (HSFb).
METHODS: Human hypertrophic scar fibroblasts were cultured in vitro, 5 cell samples were stimulated with TGF-beta 1 (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-beta 1 group (with addition of 10 ng/mL TGF-beta 1 into culture medium), CTGF ASODN group (with addition of 10% FBS-DMEM after transfection of CTGF ASODN), CTGF ASODN + TGF-beta 1group (with addition of 10 ng/mL TGF-beta 1 after transfection of CTGF ASODN). Expression of CTGF was determined by Western blotting with stimulation of different concentration of TGF-beta 1. Expression of alpha-smooth muscle actin (alpha-SMA) was measured by Western blotting. Positive cell rate of alpha-SMA was examined by flow cytometry.
RESULTS: With stimulation of 10.0 ng/mL TGF-beta 1, the expression of CTGF was obviously higher than that of non-stimulation (P < 0.05). Expression of alpha-SMA in the CTGF group and the TGF-beta 1 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-beta 1 groups (P > 0.05). The positive cell rate of alpha-SMA in NC, CTGF, TGF-beta 1, CTGF ASODN, CTGF ASODN + TGF-beta 1 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 alpha-SMA on statistic analysis.
CONCLUSIONS: CTGF is one of the most important downstream efforts for TGF-beta 1 in inducing the transdifferentiation of HSFb.
METHODS: Human hypertrophic scar fibroblasts were cultured in vitro, 5 cell samples were stimulated with TGF-beta 1 (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-beta 1 group (with addition of 10 ng/mL TGF-beta 1 into culture medium), CTGF ASODN group (with addition of 10% FBS-DMEM after transfection of CTGF ASODN), CTGF ASODN + TGF-beta 1group (with addition of 10 ng/mL TGF-beta 1 after transfection of CTGF ASODN). Expression of CTGF was determined by Western blotting with stimulation of different concentration of TGF-beta 1. Expression of alpha-smooth muscle actin (alpha-SMA) was measured by Western blotting. Positive cell rate of alpha-SMA was examined by flow cytometry.
RESULTS: With stimulation of 10.0 ng/mL TGF-beta 1, the expression of CTGF was obviously higher than that of non-stimulation (P < 0.05). Expression of alpha-SMA in the CTGF group and the TGF-beta 1 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-beta 1 groups (P > 0.05). The positive cell rate of alpha-SMA in NC, CTGF, TGF-beta 1, CTGF ASODN, CTGF ASODN + TGF-beta 1 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 alpha-SMA on statistic analysis.
CONCLUSIONS: CTGF is one of the most important downstream efforts for TGF-beta 1 in inducing the transdifferentiation of HSFb.