贵州医科大学学报

2018, v.43;No.213(06) 661-666

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microRNA-221对MPP~+诱导的帕金森病模型细胞凋亡和自噬的影响
Effect of miR-221 Expression on Apoptosis and Autophagy of Cell Modal in Patients with Parkinson's Disease Induced by MPP~+

周巍;刘鹏;刘一民;孙瑛;
ZHOU Wei;LIU Peng;LIU Yimin;SUN Ying;Department of Neurology,Xinwen Mining Group Central Hospital;Department of Neurology,Xinwen Mining Group Huayuan hospital;Department of Neurology,Yidu Central Hospital Affiliated to Weifang Medical University;

摘要(Abstract):

目的:探讨microRNA-221对1-甲基-4-苯基吡啶离子(MPP+)诱导的帕金森病(PD)模型细胞凋亡和自噬的影响。方法:用不同浓度(0.5、1和2 mmol/L)MPP+诱导SH-SY5Y细胞建立PD细胞模型,采用CCK-8试剂盒检测SH-SY5Y细胞的存活率,qRT-PCR检测不同浓度MPP+诱导建立的PD细胞模型中microRNA-221的表达;将microRNA-221 mimics及mimics-NC转染至SH-SY5Y细胞,转染后细胞分为MPP+组、MPP++mimics-NC组和MPP++microRNA-221 mimics组,qRT-PCR检测转染效果,流式细胞仪分析microRNA-221对PD细胞模型凋亡的影响,Western blotting检测microRNA-221对PD细胞模型自噬蛋白标记物(LC3II、LC3I、Beclin 1)表达的影响。结果:MPP+处理后的SH-SY5Y神经母细胞瘤细胞的细胞存活率明显降低,且MPP+对SH-SY5Y细胞的毒性呈浓度依赖性,随着MPP+浓度的增加,细胞存活率明显降低(P<0.05);随着MPP+浓度升高,PD细胞模型中microRNA-221表达水平逐渐降低(P<0.05);上调PD细胞模型中microRNA-221表达,能明显降低LC3II/LC3I比值、Beclin1蛋白表达和细胞凋亡率(P<0.05)。结论:microRNA-221在PD细胞模型中表达降低,上调microRNA-221表达能明显抑制PD细胞模型的凋亡和自噬。
Objective: To investigate the effect of microRNA-221 on apoptosis and autophagy in Parkinson's disease( PD) cell model induced by 1-methyl-4-phenylpyridine ions( MPP+). Methods:MPPMPP+at different concentrations( 0. 5、1 and 2 mmol/L) were used to induce SH-SY5 Y cell to construct the PD cell model and the survival rate of SH-SY5 Y cell was tested by CCK-8 kit. MicroRNA-221 expression levels in PD cell model induced by MPPMPP+at different concentrations were detected by qRT-PCR. MicroRNA-221 mimics and mimics-NC were transfected to SH-SY5 Y cell which was divided into group MPP、MPP++ mimics-NC and MPP++ microRNA-221 mimics after transfection. The effect of transfection was detected by qRT-PCR,the effect of microRNA-221 on PD cell model apoptosis was analyzed by using cytometer while the effect of microRNA-221 on the expression of autophagy marker in PD cell model was analyzed by using Western blotting. Results: The survival rate of SH-SY5 Y neuroblastoma cell treated by MPP+was decreased distinctly( P < 0. 05); MicroRNA-221 expression level in PD cell model was gradually decreased with the increase of MPP+concentration( P< 0. 05). Up-regulating microRNA-221 expression in PD cell model could significantly reduce the ratio of LC3 II/LC3 I and the expression of Beclin1 protein and apoptosis rate( P < 0. 05). Conclusion:Up-regulating microRNA-221 expression can apparently suppress apoptosis and autophagy of PD cell model when microRNA-221 expression in PD cell model has been down-regulated.

关键词(KeyWords): 帕金森病;细胞凋亡;自噬;1-甲基-4-苯基吡啶离子;microRNA-221
Parkinson's Disease;apoptosis;autophagy;1-methyl-4-phenylpyridine ions;microRNA-221

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作者(Author): 周巍;刘鹏;刘一民;孙瑛;
ZHOU Wei;LIU Peng;LIU Yimin;SUN Ying;Department of Neurology,Xinwen Mining Group Central Hospital;Department of Neurology,Xinwen Mining Group Huayuan hospital;Department of Neurology,Yidu Central Hospital Affiliated to Weifang Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2018.06.008

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