长链非编码RNA细胞骨架调节RNA靶向微小RNA-1246对帕金森病模型细胞损伤的保护作用

Protective effect of long non-coding RNA cytoskeleton regulatory RNA targeting microRNA-1246 on cell damage in Parkinson’s disease models

目的探讨长链非编码RNA(lncRNA)细胞骨架调节RNA(CYTOR)靶向微小RNA(miR)-1246对帕金森病(PD)模型细胞损伤的影响。方法采用100μmol/L 1-甲基-4-苯基-吡啶离子(MPP^(+))诱导SK-N-SH细胞建立PD细胞模型。Real-time PCR检测CYTOR和miR-1246表达。在SK-N-SH细胞中转染pcDNA-CYTOR或anti-miR-1246,或共转染pcDNA-CYTOR和miR-1246模拟物,经MPP^(+)处理后,采用流式细胞术分析细胞凋亡,试剂盒检测丙二醛(MDA)含量及谷胱甘肽过氧化物酶(GSH)活性。双荧光素酶报告基因实验分析CYTOR和miR-1246靶向关系。结果PD细胞模型中CYTOR表达量降低(P<0.05),miR-1246表达量升高(P<0.05)。过表达CYTOR或抑制miR-1246表达均可降低模型细胞凋亡率和MDA含量(P<0.05),并增加GSH活性(P<0.05)。MiR-1246是CYTOR的靶基因,CYTOR负性调控miR-1246表达。上调miR-1246可逆转CYTOR过表达对模型细胞凋亡和氧化损伤的影响(P<0.05)。结论LncRNA CYTOR靶向miR-1246可减轻PD模型细胞凋亡和氧化损伤。

Objective To study the effect of long non-coding RNA(lncRNA)cytoskeleton regulatory RNA(CYTOR)targeting microRNA(miR)-1246 on cell damage in Parkinson’s disease(PD)models.Methods SK-N-SH cells were exposed to 100μmol/L 1-methyl-4-phenylpyridinium(MPP+)to establish a PD cell model in vitro.The levels of CYTOR and miR-1246 were determined by Real-time PCR.pcDNA-CYTOR or anti-miR-1246,or co-transfect pcDNA-CYTOR and miR-1246 mimics were transfected into SK-N-SH cells,repectively.After treated with MPP+,flow cytometry was used to assess cell apoptosis,and commercial kits was used to monitor malondialdehyde(MDA)content and glutathione peroxidase(GSH)activity.The interaction between CYTOR and miR-1246 was verified by dual-luciferase reporter assay.Results CYTOR reduced(P<0.05),while miR-1246 increased(P<0.05)in PD model cells.CYTOR overexpression or miR-1246 inhibition could reduce the apoptosis rate and MDA content of model cells(P<0.05),and increase GSH activity(P<0.05).MiR-1246 was the target gene of CYTOR,and CYTOR negatively regulated miR-1246 expression.Up-regulation of miR-1246 could reverse the effect of CYTOR overexpression on apoptosis and oxidative damage of model cells(P<0.05).Conclusion LncRNA CYTOR can reduce cell apoptosis and oxidative damage in PD models by targeting miR-1246.