小干扰RNA抑制Vimentin表达对反应性星形胶质细胞的影响及其在胶质瘢痕形成中的意义

Influence of small interfering RNA-mediated inhibiting Vimentin on reactive astrogliosis and its effect in formation of glial scar

目的 观察小干扰RNA （siRNA）干扰波形蛋白（Vimentin）表达对反应性星形胶质细胞的影响并探讨其在中枢神经系统损伤后胶质瘢痕形成中的意义.方法 利用siRNA干扰技术沉默Vimentin表达,转染“#”字法激活的星型胶质细胞,并利用Western blot分析寻找最有效的Vimentin沉默靶点,并采用噻唑蓝（MTT）法分别于24、48、72、96 h检测反应性星形胶质细胞.结果 转染效率最高的siRNA-Vim为24 h,细胞转染效率＞80％;3条siRNA中siRNA-Vim-3对Vimentin的表达有明显的沉默效应.星形胶质细胞转染48 h后,siRNA-Vim组细胞数明显少于对照组（P＜0.05）.siRNA-Vim作用于反应性星形胶质细胞48 h后,反应性星形胶质细胞增殖受到明显抑制,与其他各组比较差异有统计学意义（P＜0.05）.结论 Vimentin对体外实验中的反应性星形胶质细胞增殖有显著的抑制作用,说明其在中枢神经系统损伤后的胶质瘢痕形成过程中起重要作用.

Objective To explore the influence of small interfering RNA （siRNA）-mediated inhibiting Vimentin on reactive astrogliosis and its effect in formation of glial scar during central nervous system injury.Methods Three different siRNA were designed and transfected astrogliosis,furthmore,Vimentin expression was silenced.We used Western blotting to search the most effective silence target.Reactive astrogliosis were tested by methyl thiazol tetrazolium （MTT） at 24,48,72 and 96 h.Results The transfection efficiency of siRNA-Vim was more than 80% when 24 h.siRNA-Vim-3 was the most efficient fragment that silenced Vimentin.Astrogliosis in siRNA-Vim group was obviously less than that of control group after transfection 48 h later （P 〈 0.05）.Reactive astrogliosis were acted by siRNA-Vim 48 h later,its proliferation was Inhibited significantly.There are significant differences compared with other groups （P 〈 0.05）.Conclusion siRNA could decrease the expression of Vimentin and inhibit the proliferation of rat astrogliosis in vitro.It was showed that Vimentin plays an important role in formation of glial scar during central nervous system injury.