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Interference of Y-27632 on the signal transduction of transforming growth factor beta type 1 in ocular tenon capsule fibroblasts

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International Journal of Ophthalmology(English edition) 2012年第5期5卷 576-581页

作者： Xiao-Hui Zhang, Jian-Ming Wang Department of Ophthalmology the Second Affiliated Hospital College of Medicine Xi'an Jiaotong University Xi'an 710004 Shaanxi Province China Department of Pharmacology Xijing Hospital Fourth Military Medical University Xi'an 710032 Shaanxi Province China

AIM: To investigate the interfering effect of Y-27632, a ROCK-I selective inhibitor, on the signal transduction pathway of transforming growth factor-beta 1 (TGF-beta 1) in ocular tenon capsule fibroblasts (OTFs) in vitro. METHODs: After OTFs from passages 4 to 6 47 vitro were induced by TGF-beta 1 and then treated by Y-27632, the changes of the OTFs cell cycles were analyzed via flow cytometry, and the proteins expression of the alpha -smooth muscular actin (alpha -sMA), connective tissue growth factor (CTGF), collagen I were calculated by Western blot. After OTFs treated by the different concentrations of Y-27632, the expression levels of the alpha -sMA, CTGF and collagen I mRNA were assayed by RT-PCR. REsULTs: Y-27632 had no markedly effect on the OTFs cell cycles. After treated by TGF-beta 1, OTFs in G1 period significantly increased. The cell cycles distribution by both TGF-beta 1 and Y-27632 had no remarkable difference from that in control group. Y-27632 significantly inhibited the proteins expressions of both alpha -sMA and CTGF, while to some extent inhibited that of collagen I. TGF-beta 1 significantly promoted the proteins expressions of alpha -sMA, CTGF and collagen I. After OTFs treated by both TGF-beta 1 and Y-27632, of alpha -sMA, the protein expression was similar with that in control group (P=0.066>0.05), but the protein expression of CTGF or collagen I, respectively, was significantly different from that in control group (P=0.000s of expressions of the alpha -sMA, CTGF and collagen I mRNA in 30, 150, 750 mu mol/L Y-27632 group were statistically significant, compared with those in control group, respectively (alpha -sMA, P=0.002, 0.000, 0.000; CTGF, P=0.014, 0.002, 0.001; collagen I,P=0.003, 0.002, 0.000). CONCLUsION: Blocking the Rho/ROCK signaling pathway by using of Y-27632 could inhibit the cellular proliferation and the expression of both CTGF and alpha -sMA whatever OTFs induced by TGF-beta 1 or not. Y-27632 suppressed t

关键词： Y-27632 ocular tenon's capsule fibroblasts transforming growth factor beta type 1 α-smooth muscular actin connective tissue growth factor collagen I

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