title: [The effect of RhoA/Rho kinase signal pathway on TGF-beta1-induced phenotypic differentiation of human dermal fibroblasts].
abstract: OBJECTIVE
  To examine the effect of RhoA/Rho kinase signal pathway on TGF-beta1-induced phenotypic differentiation of human dermal fibroblasts.
  
  
  METHODS
  The 4th generation of primary cultured human dermal fibroblasts were stimulated with TGF-beta1, (10 ng/ml). The expression of alpha-SMA was detected after treatment with TGF-beta1, for 0, 3, 6, and 24 h. The expression of alpha-SMA was also detected after treatment with different concentration of TGF-beta1 (0, 2, 10, 50 ng/ml). Then the human dermal fibroblasts (4th generation) were stimulated with TGF-beta1, (10 ng/ml) after being treated with the RhoA/Rho kinase signaling pathway inhibitor Y-27632 (10 umol/ml). The fibroblasts were treated with nothing as sham control, or with Y-27632 (10 umol/L) only as negative control group, or with TGF-beta1 (10 ng/ml) only as positive control group. The expression of alpha-SMA was detected in all the groups. Protein expression was analyzed with ANOVA statistical method.
  
  
  RESULTS
  alpha-SMA expression in fibroblasts with 10 ng/ml TGF-beta1 stimulation for 0, 3, 6, 24 h was 1.0, 1.9 0.2, 2.1 +/- 0. 1, 3. 1 +/- 0.1, respectively. Alpha-SMA expression in 24 h group was significantly higher than that in other three groups (n = 4, P < 0.05). alpha-SMA expression in human dermal fibroblasts after stimulation with different concentration of TGF-beta1 (0, 2, 10, 50 ng/ml) was 1.0, 1.4 +/- 0.2, 3.2 + 0.1, 3.1 +/- 0.2, respectively. alpha-SMA expression in 10 ng/ ml group was significantly higher than that in 2 ng/ml group and control group (n = 4, P < 0.05). There was no statistical difference in alpha-SMA expression between 10 ng/ml group and 50 ng/ml group (n = 4, P > 0.05). With both Y-27632 (10 micromol/L) and TGF-beta1 stimulation, the cell phenotype differentiation was inhibited. Alpha-SMA expression in experimental group (1.2 +/- 0.2) was significantly reduced, when compared with that in positive control group (2.9 +/- 0.1) (n = 5, P < 0.05). There was no significant difference (n = 5, P > 0.05) in alpha-SMA expression between control group (1.0) and negative control group (1.1 +/- 0.1).
  
  
  CONCLUSIONS
  RhoA/Rho kinase signaling pathway should be involved in TGF-beta1-induced phenotypic differentiation of human dermal fibroblasts.
authors.affiliation: Center of Plastic Surgery and Burn Repair, 309th Hospital of PLA, Beijin 100091, China.
authors.forename: Yong-Ling
authors.initials: YL
authors.lastname: Hu
authors.forename: Zhen
authors.initials: Z
authors.lastname: Liu
authors.forename: Da-Kai
authors.initials: DK
authors.lastname: Jiao
authors.forename: Tian
authors.initials: T
authors.lastname: Ma
authors.forename: Chang-Yang
authors.initials: CY
authors.lastname: Wang
authors.forename: Chi-Yu
authors.initials: CY
authors.lastname: Jia
mesh_terms: D000199:Actins / Q000378:metabolism; D000293:Adolescent; D002454:Cell Differentiation; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D008297:Male; D015398:Signal Transduction; D012867:Skin / Q000166:cytology; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*; D054460:rho-Associated Kinases / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
chemical_list: D000199:Actins; D053773:Transforming Growth Factor beta1; C066737:RHOA protein, human; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein