中文摘要：

目的研究ski基因在大鼠星形胶质细胞迁移过程中的作用。方法分离大鼠脑皮质星形胶质细胞,体外培养。合成ski基因和阴性对照小干扰片段。设置ski-siRNA转染组、阴性对照组和空白对照组。采用脂质体法将特异性针对ski基因的siRNA和阴性对照siRNA转入大鼠星形胶质细胞。采用Western blotting检测各组ski蛋白的表达水平;采用Transwell细胞迁移实验和细胞划痕实验检测ski-siRNA转染后星形胶质细胞迁移能力的变化。结果与空白对照组和阴性对照组比较,ski-siRNA转染组细胞内ski蛋白的相对表达水平显著降低（F=132.957,P〈0.001）。Transwell细胞迁移实验发现,ski-siRNA转染组每孔迁移细胞数少于空白对照组和阴性对照组（F〉47.197,P〈0.05）。细胞划痕实验显示,转染组细胞1~5 d的划痕愈合率均明显低于对照组（F〉69.187,P〈0.01）。结论沉默ski基因能显著抑制星形胶质细胞的迁移能力,ski可能参与星形胶质细胞的转移过程,进而调控胶质瘢痕的形成。

英文摘要：

Objective To investigate the effect of ski gene in migration process of astrocytes in rats.Methods Astrocytes were obtained from rats＇ cerebral cortex and cultured in vitro.siRNA targeting ski gene and negative control sequences were prepared.The ski-siRNA group,siRNA negative control group and untreated group were set in this experiment.The specific siRNA targeting ski gene was transfected into astrocytes with Lipofectamine~？RNAi MAX Reagent.Then the ski protein levels were determined with Western blotting.After transfection,the changes in migration of astrocytes were measured with wound scratch assay and Transwell migration assay.Results Western blotting showed that the expression of ski protein was significantly lower in the ski-siRNA group than in the siRNA negative control group and untreated group（F=132.957,P〈0.001）.Transwell migration assay showed that the number of astrocytes crossing through chambers was less in the ski-siRNA group than in the siRNA negative control group and untreated group（F〉47.197,P〈0.05）.Wound scratch assay showed that the wound healing rate was lower in the ski-siRNA group than in the control group one,two,three,four and five days after transfection（F69.187,P〈0.001）.Conclusion Ski knocked down by siRNA could inhibit the migration ability of astrocytes.It is a reminding that ski may take part in the migration process of astrocytes,and moreover,ski may play an important role in the formation of glial scar.