ROCK-Ⅰ选择性抑制剂Y-27632对人Tenon囊成纤维细胞增殖、凋亡和黏附性的影响

Effect of Y-27632,a selective inhibitor of ROCK-I,on proliferation,apoptosis and adhesion of human ocular Tenon's capsular fibroblasts

目的观察ROCK-Ⅰ选择性抑制剂Y-27632对体外培养的人眼Tenon囊成纤维细胞(ocular Tenon capsule fibroblasts,OTFs)增殖、凋亡和黏附性的影响。方法体外培养的第4～6代OTFs经溶血磷脂酸(lysophosphatidic acid,LPA)诱导后,不同浓度Y-27632处理,采用MTT测定细胞增殖抑制率和细胞黏附抑制率,未经LPA诱导OTFs细胞同时采用Annexin V-FITC/PI双标记流式细胞术测定细胞凋亡率,以及细胞平板克隆增殖实验测定不同浓度Y-27632组OTFs细胞增殖克隆形成率。结果 6、30、150、750μmol/L Y-27632各组经LPA诱导的OTFs细胞增殖抑制D(490)与LPA组比较,差异均有统计学意义(P<0.05,P<0.01),OTFs细胞黏附抑制D(490)与LPA组比较,同样具有统计学差异(P<0.05,P<0.01)。随着Y-27632浓度增加,细胞凋亡率相应增加。未经LPA诱导的OTFs细胞克隆形成率随着Y27632浓度的增高而降低。结论 Y-27632有效抑制LPA诱导的OTFs增殖和细胞间黏附,诱导细胞早期凋亡。

Objective To investigate the effect of Y-27632,a selective inhibitor of ROCK-I,on the proliferation,apoptosis and adhesion of human ocular Tenon＇s capsular fibroblasts（OTFs） in vitro.Methods After induced by lysophosphatidic acid（LPA）,the fourth to sixth passages of OTFs were exposed to 6,30,150 and 750 μmol/L of Y-27632 for 48 h,respectively,and MTT assay was applied to determine the inhibition ratios of cell proliferation and adhesion.OTFs without LPA induction were treated with different concentrations of Y-27632,and Annexin V-FITC/PI double labeling flow cytometry was applied to determine cell apoptotic rate.The cell cloning efficiency was calculated by cell plate clonal expansion experiment.Results The proliferation inhibition D（490） and adhesion inhibition D（490） of OTFs treated with LPA＋Y-27632 were significantly greater than those of LPA-induced OTFs（P0.05,P0.01）.Cell apoptotic rate increased along with the increase of Y-27632 concentrations,and the cloning efficiency of OTFs without LPA induction decreased along with the increase of Y-27632 concentrations.Conclusion Y-27632 can effectively inhibit both proliferation and adhesion of LPA-induced OTFs,and induce early apoptosis of OTFs without LPA induction.