法舒地尔和RhoA沉默对雪旺细胞增殖的影响王玉成1周江波2.DOCVIP

法舒地尔和RhoA沉默对雪旺细胞增殖的影响 王玉成1 周江波2（301600 天津, 天津市静海县医院神经外科1 天津市第四中心医院神经外科2） ［提要］目的 观察Rho激酶抑制剂法舒地尔和RNAi介导的RhoA基因沉默对大鼠雪旺细胞增殖的影响。方法　体外培养Wistar大鼠雪旺细胞，分6组干预：空白对照组，5，10，15，20μmol/L 法苏地尔组，siRNA 沉默RhoA基因组。处理后第3天，采用RT-PCR，Western blot检测各组雪旺细胞RhoA基因和蛋白的表达。连续5天用细胞计数法测定生长曲线。应用MTT比色法观察μmol/L 法苏地尔组、siRNA 沉默RhoA基因组雪旺细胞RhoA基因和蛋白表达量较5，10μmol/L 法苏地尔组、空白对照组明显降低（P0.05），细胞的生长速度较5，10μmol/L 法苏地尔组、空白对照组明显降低明显增快（P0.05），细胞周期G0/G1期减少（P0.05），S期细胞数增多（P0.05）。法苏地尔浓度增加到20μmol/L时对细胞的作用并非随浓度的增加而增强，与15μmol/L组的差异无显著性意义（P﹥0.05）。15，20μmol/L 法苏地尔组与siRNA 沉默RhoA基因组相比差异相比RhoA基因和蛋白表达量有差异有统计学意义（P0.05）。结论　Rho激酶抑制剂法舒地尔和RNAi介导的RhoA基因沉默在体外实验均能促进雪旺细胞，法舒地尔最佳作用浓度为15μmol/L，沉默RhoA基因效果较好。 ［关键词］雪旺细胞; RhoA; RNA干扰;法舒地尔； ［中图法分类号］R687 ［文献标识码］ A Effects of Fasudil and RhoA silencing on the proliferation of Schwann cells. WANG yu-cheng1 ZHOU jiang-bo2 , (Department of Neurosurgery, Tianjin Jinghai Hospital1, Department of Neurosurgery, Tianjin Fourth Central Hospital2. Tianjin 301600, P.R.China.) ［Abstract］Objective To determine whether the ROCK inhibit or RhoA gene silencing in Schwann cells can optimized Schwann cells cultures . Methods Schwann cells was cultured by the suspension culture in vitro, and were divided into 5 groups: A group was culture cells with 0μmol/L Fasudil; B group was culture cells with 5μmol/L Fasudil;C group was culture cells with 10μmol/L Fasudil; D group was culture cells with 15μmol/L Fasudil;E group was culture cells with 20μmol/L Fasudil ;F group was RhoA gene silencing group .Three days later, RT-PCR and western blot were used to assess the expression of RhoA mRNA and RhoA protein. Cellular proliferation was determined by the cell growth curve (five days) and MTT assay. The cell cycle was analyzed by flow cytometry. Results three days later, D, E, F group compared with A, B, C group RhoA gene and protein expression was significantly lower (P 0.05), cell growth rate significantly increased, cell cycle G0/G1 reduce, S phase cells increased. There were significant differences among the g