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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。
Objective To investigate the effect of miR-133b on the expression of tyrosine hydroxylase (TH) in Parkinson’s disease (PD) dopaminergic neurons induced by 1-methyl-4-phenylpyridinium ion (MPP +). Methods The primary cultured cells of ventral midbrain of rat embryo (14 d pregnant) were divided into 4 groups according to different treatment methods: 1 blank control group without any intervention; 2 negative control group, The lentiviral particles (miR-NC) were transfected 48 h after transfection and the cells were cultured in serum-free medium for 24 h. The cells were transfected with miRNA lentiviral vector for 48 h in 3miR-NC + MPP + The cells were transfected with 10μmol·L-1 MPP + for 24 h. The cells were transfected with miR-133b lentivirus for 48 h. The cells were treated with 10μmol·L-1 MPP + for 24 h. Western blot was used to detect the expression of TH protein in each group. Results Compared with miR-NC + MPP + group, the expression of TH protein in miR-133b + MPP + group was significantly increased, the difference was statistically significant (F = 22.598, P <0.05). Conclusion miR-133b can enhance the expression of TH protein in PD dopaminergic neurons and play a neuroprotective role.