摘要
背景:许旺细胞在周围神经损伤和修复再生过程中发挥重要作用。目的:观察Rho激酶抑制剂法舒地尔和RNAi介导的RhoA基因沉默对大鼠许旺细胞增殖的影响。方法:体外培养Wistar大鼠许旺细胞,分6组干预: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,RT-PCR、Westernblot检测各组许旺细胞RhoA基因和蛋白的表达。连续5d用细胞计数法测定生长曲线。应用MTT比色法观察许旺细胞增殖情况;采用流式细胞术测定许旺细胞周期分布的变化。结果与结论: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)。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许旺细胞,法舒地尔最佳作用浓度为15μmol/L,沉默RhoA基因效果较好。
BACKGROUND：Schwann cells play an important role in peripheral nerve injury,regeneration and repair.OBJECTIVE：To determine whether Fasudil or RNAi-mediated RhoA gene silencing can optimize Schwann cells culture.METHODS：Schwann cells were divided into 5 groups：control group;5 μmol/L Fasudil group;10 μmol/L Fasudil group;15 μmol/L Fasudil group;20 μmol/L Fasudil group;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（5 days） and MTT assay.The cell cycle was analyzed by flow cytometry.RESULTS AND CONCLUSION：There was no difference between 20 μmol/L and 15 μmol/L Fasudil groups（P〉0.05）.Fasudil and RNAi-mediated RhoA gene silencing can promote the proliferation of Schwann cells,and 15 μmol/L Fasudil and RhoA gene silencing have the best effect.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2011年第49期9239-9243,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
