miR-15b对MPP^+损伤SH-SY5Y细胞中α突触核蛋白表达的影响

Effect of microRNA-15b on the expression of α-synuclein in SH-SY5Y cells with 1-methyl-4-phenylpyridinium-induced injury

目的研究miR-15b对MPP+(1-甲基-4-苯基-吡啶离子)损伤SH-SY5Y细胞中α突触核蛋白表达的影响以及在帕金森病(Parkinson disease,PD)发病机制中的作用。方法通过CCK-8检测出MPP+诱导SH-SY5Y细胞为PD细胞模型的最适浓度和时间,然后将过表达和沉默miR-15b的质粒转染到PD细胞模型内,再通过Real Time-PCR和Western Blot检测miR-15b和α突触核蛋白的表达量。结果 SH-SY5Y细胞经MPP+诱导后细胞形态发生改变、细胞增殖能力降低,过表达miR-15b后α-synuclein和α突触核蛋白的表达量均降低(P <0. 05),沉默miR-15b后α-synuclein和α突触核蛋白的表达量均升高(P <0. 05)。结论 miR-15b可以抑制PD细胞模型内α-synuclein和α突触核蛋白的表达。miR-15b可能参与了帕金森病患者中脑黑质多巴胺能神经元内α突触核蛋白的富集调节机制。

Objective To investigate the effect of microRNA-15b( miR-15b) on the expression of α-synuclein in SH-SY5 Y cells with1-methyl-4-phenylpyridinium( MPP^+)-induced injury and its role in the pathogenesis of Parkinson’s disease(PD). Methods CCK-8 was used to determine the optimal concentration and time of MPP+induction in the establishment of a PD cell model using SH-SY5 Y cells,and then the PD cell model was transfected with the plasmids with overexpressed or silenced miR-15 b. Real-time PCR and Western blot were used to measure the mRNA and protein expression of miR-15 b and α-synuclein. Results SH-SY5 Y cells had changes in cell morphology and a reduction in cell proliferation after MPP^+ induction. There were significant reductions in the mRNA and protein expression of α-synuclein after miR-15 b overexpression( P < 0. 05),and there were significant increases in the mRNA and protein expression of α-synuclein after miR-15 b silencing( P < 0. 05). Conclusions MiR-15 b can inhibit the mRNA and protein expression ofα-synuclein in the PD cell model. MiR-15 b may be involved in the enrichment and regulation of α-synuclein in the substantia nigra dopaminergic neurons in patients with PD.