LncRNA MAGI2-AS3靶向调控miR-1277-5p对帕金森病细胞模型损伤的影响

The effect of LncRNA MAGI2-AS3 on the injury of Parkinson’s disease cell model through targeted regulation of miR-1277-5p

目的探讨LncRNA MAGI2-AS3对帕金森病细胞模型损伤的影响及其对miR-1277-5p的调控作用。方法用1-甲基-4-苯基自由基离子(MPP+)诱导的SK-N-SH细胞建立帕金森病细胞模型,实验分组:MPP++si-NC组、MPP++si-MAGI2-AS3组、MPP++miR-NC组、MPP++miR-1277-5p组、MPP++si-MAGI2-AS3+anti-miR-NC组、MPP++si-MAGI2-AS3+anti-miR-1277-5p组;qRT-PCR检测MAGI2-AS3与miR-1277-5p的表达量;根据试剂盒检测LDH、MDA、GSH的含量;CCK-8、流式细胞术分别检测细胞增殖及凋亡;双荧光素酶报告基因实验检测MAGI2-AS3与miR-1277-5p的靶向关系。结果MPP+诱导的SK-N-SH细胞中MAGI2-AS3表达量升高(P<0.05),miR-1277-5p表达量降低(P<0.05),LDH、MDA水平和凋亡率升高(P<0.05),细胞存活率和GSH水平降低(P<0.05);转染si-MAGI2-AS3或转染miR-1277-5p mimics可抑制MPP+诱导的SK-N-SH细胞凋亡及氧化应激,并可促进细胞增殖;MAGI2-AS3可靶向调控miR-1277-5p;共转染si-MAGI2-AS3与anti-miR-1277-5p能够逆转干扰MAGI2-AS3表达对MPP+诱导的SK-N-SH细胞氧化应激、增殖及凋亡作用。结论干扰MAGI2-AS3表达可通过上调miR-1277-5p而抑制SK-N-SH细胞氧化应激反应、凋亡及促进细胞增殖进而减轻帕金森病细胞模型损伤。

Objective To explore the effect of LncRNA MAGI2-AS3 on the damage of Parkinson’s disease cell model and its regulatory effect on miR-1277-5p.Methods 1-methyl-4-phenyl radical ion(MPP+)was used to induce SK-N-SH cells to establish a Parkinson’s disease cell model.Experimental groups were divided as follows:MPP++si-NC group,MPP++si-MAGI2-AS3 group,MPP++miR-NC group,MPP++miR-1277-5p group,MPP++si-MAGI2-AS3+anti-miR-NC group and MPP++si-MAGI2-AS3+anti-miR-1277-5p group,respectively.qRT-PCR assay was used to detect the expression of MAGI2-AS3 and miR-1277-5p.The contents of LDH,MDA and GSH were tested according to the kit instruction.CCK-8 and flow cytometry were used to detect cell proliferation and apoptosis.The dual luciferase reporter gene assay was used to detect the targeting relationship between MAGI2-AS3 and miR-1277-5p.Results The expression of MAGI2-AS3 in SK-N-SH cells induced by MPP+was increased(P<0.05),accompanied with decreased expression of miR-1277-5p(P<0.05).Meanwhile,the level of LDH,MDA and the rate of apoptosis were increased significantly(P<0.05);Conversely,cell survival rate and the level of GSH were decreased obviously(P<0.05).Transfection of si-MAGI2-AS3 or miR-1277-5p mimics could inhibit MPP+-induced SK-N-SH cell apoptosis and oxidative stress,and can promote cell proliferation.MAGI2-AS3 could target miR-1277-5p.Co-transfection of si-MAGI2-AS3 and anti-miR-1277-5p could reverse the effect of interference with the expression of MAGI2-AS3 on the oxidative stress,proliferation and apoptosis of SK-N-SH cells induced by MPP+.Conclusion Interfering with the expression of MAGI2-AS3 could inhibit the oxidative stress response,apoptosis and promote cell proliferation of SK-N-SH cells by up-regulating miR-1277-5p,thereby reducing the damage of Parkinson’s disease cell models.