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

作　　者：谢时帅 牛万祥 徐峰 胡珊珊 牛朝诗 XIE Shi-shuai;NIU Wan-xiang;XU Feng;HU Shan-shan;NIU Chao-shi(Department of Neurosurgery,Anhui Provincial Hospital Affiliated to Anhui Medical University,Anhui Provincial Key Laboratory of Brain Function and Brain Disease,Hefei 230001,China)

机构地区：[1]安徽医科大学附属省立医院神经外科,安徽合肥230001 [2]脑功能与脑疾病安徽省重点实验室,安徽合肥230001 [3]安徽省脑立体定向神经外科研究所,安徽合肥230001 [4]安徽省神经系统疾病(神经外科)临床研究中心,安徽合肥230001

出　　处：《国际神经病学神经外科学杂志》2018年第6期591-596,共6页Journal of International Neurology and Neurosurgery

基　　金：中央引导地方科技发展专项(编号:2017070802D144);脑功能与脑疾病安徽省重点实验室绩效考核补助项目(编号:1606c08235);国家重点研发计划项目子课题(编号:2016YFC0105900)

摘　　要：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.

关 键 词：帕金森病 miR-15b MPP+ SH-SY5Y Α突触核蛋白 

分 类 号：R742.5[医药卫生—神经病学与精神病学]