| 李文娟,聂尚丹,亚白柳,王春梅.miR let-7f对流行性乙型脑炎病毒在神经元细胞中复制的调控作用[J].济宁医学院学报,2019,42(6):390-394 |
| miR let-7f对流行性乙型脑炎病毒在神经元细胞中复制的调控作用 |
| miR let-7f inhibited Japanese encephalitis virus replication in neurons |
| 投稿时间：2019-03-26 |
| DOI：10.3969/j.issn.1000-9760.2019.06.003 |
| 中文关键词: 流行性乙型脑炎病毒;miR let-7f;神经元;SH-SY5Y细胞 |
| 英文关键词: Japanese encephalitis virus;miR let-7f;Neuron;SH-SY5Y cells |
| 基金项目:山东省自然科学基金（ZR2017LH056，ZR2015PH026）；国家自然科学基金（81501018，81703490） |
| 作者 | 单位 | E-mail | | 李文娟 | 济宁医学院法医学与医学检验学院, 济宁 272067 | | | 聂尚丹 | 济宁医学院法医学与医学检验学院, 济宁 272067 | | | 亚白柳 | 济宁医学院基础医学院, 济宁 272067 | | | 王春梅 | 济宁医学院神经生物学研究所, 济宁 272067 | wangchunmei410@mail.jnmc.edu.cn |
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| 中文摘要: |
| 目的 探讨miR let-7f对流行性乙型脑炎病毒（Japanese encephalitis virus，JEV）在神经元细胞中复制的影响。方法 培养人神经母细胞瘤SH-SY5Y细胞，miRNA芯片检测SH-SY5Y细胞感染JEV后miRNA表达谱，荧光定量RT-PCR检测确认miR let-7f表达情况；实验随机分为miR let-7f mimics组、miR let-7f inhibitor组和miRNA conrol组，分别将miR let-7f mimics、inhibitors和miRNA conrol转染SH-SY5Y细胞后感染JEV，荧光定量RT-PCR和空斑法检测病毒复制变化情况。结果 SH-SY5Y细胞感染JEV后差异表达的miRNA有37个，上调的有28个，下调的有9个，其中miR let-7f表达明显升高（3.51倍）；SH-SY5Y细胞感染JEV 12h后miR let-7f表达明显上升（3.67倍），与miRNA control组和miR let-7f inhibitor组相比，miR let-7f mimics组细胞培养上清中病毒滴度降低，细胞内JEV RNA也显著减少，差异具有统计学意义（P<0.025）。结论 miR let-7f在神经元细胞中能够有效抑制JEV复制，在开发JEV感染的分子治疗方面具有潜在应用价值。 |
| 英文摘要: |
| Objective To analyze the role of miR let-7f on Japanese encephalitis virus (JEV) infection and replication in neurons.Methods SH-SY5Y cells were cultured,and miRNA chip was used to detect differentially expressed miRNAs between SH-SY5Y cells after JEV infection and normal cells.The relative expression level of miR let-7f related to JEV infection was determined by qRT-PCR.SH-SY5Y cells were divided into three groups randomly which transfected with miR let-7f mimics,inhibitors,and miRNA conrol respectively,and then infected with JEV.Infected cells and supernatant were harvested and then qRT-PCR and plaque forming test were used to detect the levels of virus replication.Results A total of 28 miRNAs were up-regulated while 9 miRNAs down-regulated in SH-SY5Y cells infected with JEV compared with non-infected cells,of which miR let-7f up-regulated 3.51 folds.miR let-7f was confirmed to be up expressed 3.67 folds in SH-SY5Y cells after JEV infection 12h.After miR let-7f mimics were transfected,JEV RNA in cells and virus titer in supernatant was lower than miR let-7f inhibitors/miRNA conrol groups,and the differences showed statically significant (P<0.025).Conclusion miR let-7f could inhibit JEV replication in neurons which might be as a potential molecular target for novel therapy. |
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