摘要
目的探讨miR-133b对l-甲基-4-苯基吡啶离子(MPP+)诱导的帕金森病(PD)多巴胺能神经元模型中酪氨酸羟化酶(TH)表达量的影响。方法采用大鼠胚胎(孕14 d)中脑腹侧组织进行原代细胞培养,依据不同处理方法分为4组:1空白对照组,无任何干预措施;2阴性对照组,使用携带空载miRNA慢病毒颗粒(miR-NC)持续转染48 h,换液后无血清培养基继续培养24 h;3miR-NC+MPP+组,使用携带空载miRNA慢病毒颗粒持续转染48 h,换液后10μmol·L-1MPP+继续作用24 h;4miR-133b+MPP+组,使用携带miR-133b慢病毒颗粒持续转染48 h,换液后10μmol·L-1 MPP+继续作用24 h。采用Western blot法分别检测各组TH蛋白表达水平。结果与miR-NC+MPP+组比较,miR-133b+MPP+组TH蛋白表达量显著升高,差异有统计学意义(F=22.598,P<0.05)。结论 miR-133b可提高PD多巴胺能神经元模型中TH蛋白表达水平,发挥一定的神经保护作用。
Aim To evaluate the effect of miR-133 b on expression of tyrosine tydroxylase（TH） in primary cell model of 1-methyl-4-phenyl-pyridinium（MPP＋） induced Parkinson＇s disease（PD）. Methods Embryonic rat mesencephalic primary neurons were cultured in vitro and divided into four groups： a blank control group, untreated cells; a negative control group,neurons were transfected with lentivirus containing negative control miRNA（miR-NC） for 48 h; a miR-NC＋MPP＋ group, neurons were injured by MPP＋ for 24 h and pretreated with miR-NC for 48 h; a miR-133 b ＋MPP＋ group, neurons were transfected with lentivirus containing miR-133 b for 48 h before injured by MPP＋ for another 24 h. The protein levels of TH was measured by Western blot. Results Compared with MPP＋group, the level of TH protein significant Iy increased in miR-133 b group. Conclusion The results suggested that overexpression of miR-133 b may play an important role in protecting dopaminergic neurons.
出处
《中国临床神经科学》
2016年第2期133-138,共6页
Chinese Journal of Clinical Neurosciences
基金
国家自然科学基金资助项目(编号:81571225)
