siRNA沉默卫星胶质细胞轴突导向因子Slit1对共培养的神经元突起生长的影响  

Effect of siRNA-mediated gene silencing of Slit1 in satellite glial cells on growth of neurite

作　　者：张彩彩[1,2] 赵久红[2,3] 谭晓红[2] 易西南[2,3] ZHANG Cai-cai;ZHAO Jiu-hong;TAN Xiao-hong;YI Xi-nan(Department of Physiology 1.Laboratory for Neuroscience;Department of Anatomy;Hainan Medical College,Haikou 571199,Hainan,CHINA)

机构地区：[1]海南医学院生理教研室,海南海口571199 [2]海南医学院神经科学实验室,海南海口571199 [3]海南医学院解剖教研室,海南海口571199

出　　处：《海南医学》2018年第17期2369-2372,共4页Hainan Medical Journal

基　　金：海南省自然科学基金(编号:20158308);海南医学院培育基金(编号:HY2014-006)

摘　　要：目的观察胶质细胞Slit1对脊髓背根节(DRG)神经元突起生长的影响。方法构建大鼠Slit1siRNA,转染至大鼠原代DRG卫星胶质细胞。利用实时荧光定量-聚合酶链反应(RT-PCR)、Western blot技术检测Slit1 m RNA和蛋白的表达情况;将siRNA-Slit1转染成功的胶质细胞与大鼠原代DRG神经元共培养48 h,显微镜下观察神经元突起生长情况。结果 (1)与阴性对照组比较,siRNA-Slit1组胶质细胞Slit1 m RNA的表达量明显降低,差异具有统计学意义(P<0.05);空白对照组与阴性对照组比较差异无统计学意义(P>0.05);(2)siRNA-Slit1组Slit1蛋白的表达量明显低于阴性对照组,差异具有显著统计学意义(P<0.01),而空白对照组与阴性对照组间比较差异无统计学意义(P>0.05);(3)siRNA-Slit1组神经元突起长度明显短于阴性对照组,差异具有显著统计学意义(P<0.01)。结论 Slit1是诱导神经元突起生长的重要导向分子,可促进离体DRG神经元突起的生长。Objective To investigate the effect of Slitl in satellite glial cells on neurite outgrowth of dorsal root ganglion （DRG） neurons. Methods The rat Slitl siRNA was constructed and transfected into rat primary DRG glial cells. Real time fluorescence quantitative polymerase chain reaction （RT-PCR） and Western blot were used to detect the expression of Slit1 rnRNA and protein. The glial cells transfected with siRNA-Slitl were co-cultured with primary DRG neurons for 48 h, and the neurite outgrowth was observed under microscope. Results （1） RT-PCR results showed that Slit1 mRNA expression in glial cells was significantly lower in siRNA-Slitl group than negative control group （P〈 0.05）. There were no significant differences in Slitl mRNA expression between the blank control group and the negative control group （P〉0.05）. （2） The results of western blot showed that the expression of Slitl protein in the glial cells was significantly lower than that in negative control group （P〈0.01）. There were no significant differences between the blank control group and the negative control group （p〉0.05）. （3） The neurite length in siRNA-Slitl group was significantly shorter than that in negative control group （P〈0.01）. Conclusion Slitl protein is an important guide molecule for neurite outgrowth, and it promotes nettrite outgrowth in vitro.

关 键 词：Slit1 胶质细胞 神经元 基因沉默 共培养 

分 类 号：R741[医药卫生—神经病学与精神病学]