miRNA-145通过下调ROCK1的表达抑制人牙周膜成纤维细胞的迁移

miRNA-145 Inhibits the Migration of Human Periodontal Ligament Fibroblasts via Down-regulating the Expression of ROCK1

目的:研究微小核糖核酸-145(microRNA-145,miRNA-145)对人牙周膜成纤维细胞迁移的影响及其作用机制。方法:体外采用酶消化法培养人牙周膜成纤维细胞并传代,将其分为对照组和转染miRNA-145组,按50 ng/mL的miRNA-145浓度转染人牙周膜成纤维细胞,转染72 h后提取各组蛋白,用Western blot检测miRNA-145的靶蛋白ROCK1的表达水平的相关变化;采用划痕试验检测各组划痕细胞间距离的相关变化,选取划痕后的0 h、24 h、48 h、72 h时间点,测量各时间点划痕细胞间的距离并计算平均值。结果:与对照组相比,转染miRNA-145后,miRNA-145靶蛋白ROCK1的表达量显著降低(p<0.05);转染24 h、48 h后细胞间距离的均值大于对照组(p<0.05)。结论:miRNA-145可能通过下调ROCK1的表达抑制人牙周膜成纤维细胞的迁移。

Objective： To study the effect and mechanism of miRNA-145 on the migration of human periodontal ligament fibroblasts. Methods： The human periodontal ligament fibroblasts were cultured by the method of enzyme digestion in vitro and gone down to the future generation. The cells were divided into two groups. The cells in experimental group were transfected with 50 ng/mL miRNA-145. The protein expression of ROCK1 was analyzed by Western Blot. Scratch test was performed to measure the changes of distances between the scratched cells at 0, 24, 48, 72 hours after scratching. Results： Compared with the control group, the expression level of ROCK1 was significantly lower in the experimental group（p0.05）; and the distance between the scratched cells of experimental group was significantly higher than that of the control group at 24, 48 hours after transfection（p0.05）. Conclusions： miRNA-145 could inhibit the migration of human periodontal ligament fibroblasts via down-regulating the expression of ROCK1.