Department of Neurology, Jingzhou Central Hospital, Jingzhou, Hubei, China.
Department of Neurology, Jingzhou Central Hospital, Jingzhou, Hubei, China.
Neurosci Lett. 2019 Aug 24;708:134340. doi: 10.1016/j.neulet.2019.134340. Epub 2019 Jun 19.
Long non-coding RNAs (lncRNAs) have been reported to play important roles in Parkinson's disease (PD) pathogenesis. It was indicated that lncRNA nuclear enriched abundant transcript 1 (NEAT1) is involved in PD. However, the underlying mechanism of NEAT1 is still not fully explored. Human neuroblastoma cell line SH-SY5Y was treated with 1-Methyl-4-phenylpyridinium (MPP) to mimic PD model in vitro. qRT-PCR was employed to detect the expression of NEAT1, IL-1β, IL-6 and TNF-α. Starbase database, RNA pull-down assay and RNA immunoprecipitation (RIP) assay were performed to verify the relationship between NEAT1 and miR-124. MTT assay and flow cytometry assay were used to detect cell viability and apoptosis. Elisa was introduced to measure the levels of IL-1β, IL6 and TNF-α in culture media. We found that NEAT1 expression was significantly increased in SH-SY5Y cells after MPP treatment in dose- and time- dependent manners. MPP induced the expression and secretion of IL-1β, IL-6 and TNF-α, inhibited cell viability and induced apoptosis while these effects could be rescued by NEAT1 silencing. miR-124 was a target of NEAT1. Anti-miR-124 could reverse the effects caused by NEAT1 knockdown in MPP treated SH-SY5Y cells. Therefore, we speculated that NEAT1 may regulate MPP induced neuronal injury by targeting miR-124 in SH-SY5Y cells.
长链非编码 RNA(lncRNA)在帕金森病(PD)发病机制中发挥重要作用。有研究表明,lncRNA 核富集丰富转录物 1(NEAT1)参与 PD 的发生。然而,NEAT1 的潜在机制仍未完全阐明。用人神经母细胞瘤细胞系 SH-SY5Y 用 1-甲基-4-苯基吡啶(MPP)处理,体外模拟 PD 模型。qRT-PCR 检测 NEAT1、IL-1β、IL-6 和 TNF-α 的表达。Starbase 数据库、RNA 下拉实验和 RNA 免疫沉淀(RIP)实验验证 NEAT1 与 miR-124 之间的关系。MTT 实验和流式细胞术检测细胞活力和凋亡。Elisa 法检测培养上清液中 IL-1β、IL6 和 TNF-α 的水平。结果发现,MPP 处理呈剂量和时间依赖性增加 SH-SY5Y 细胞中 NEAT1 的表达。MPP 诱导 IL-1β、IL-6 和 TNF-α 的表达和分泌,抑制细胞活力并诱导细胞凋亡,而 NEAT1 沉默可逆转这些作用。miR-124 是 NEAT1 的靶标。抗 miR-124 可逆转 MPP 处理的 SH-SY5Y 细胞中 NEAT1 敲低引起的效应。因此,我们推测 NEAT1 可能通过靶向 SH-SY5Y 细胞中的 miR-124 来调节 MPP 诱导的神经元损伤。