Geng Lijiao, Zhao Jun, Liu Wei, Chen Yong
Department of Rehabilitation Medicine, Huaihe Hospital of Henan University No. 357 Ximen Street Kaifeng 475000 China
Department of Neurology, Huaihe Hospital of Henan University Kaifeng 475000 China.
RSC Adv. 2019 Aug 13;9(43):25257-25265. doi: 10.1039/c9ra05039f. eCollection 2019 Aug 8.
Long noncoding RNAs (lncRNAs) have recently attracted increasing attention for their involvement in a wide variety of human neurodegenerative diseases, including Parkinson's disease (PD). The purpose of the present study was to investigate the functional role and underlying mechanism of NEAT1 in PD. qRT-PCR was used to assess the expression of NEAT1 and miR-221, and the expression levels of Bcl-2 and Bax were detected by western blot. Cell viability and apoptosis were determined by CCK-8 assay and flow cytometry, respectively. The changes of oxidative stress and neuroinflammation were evaluated by ELISA assay and qRT-PCR, respectively. The targeted interaction between NEAT1 and miR-221 was verified by dual-luciferase reporter assay and RNA immunoprecipitation assay. Our data supported that MPP treatment elevated NEAT1 expression in dose- and time-dependent manners in SH-SY5Y cells, and NEAT1 silencing relieved MPP-induced suppression of cell viability and enhancement of cell apoptosis in SH-SY5Y cells. Moreover, NEAT1 silencing alleviated MPP-induced promotion of oxidative stress and neuroinflammation in SH-SY5Y cells. NEAT1 directly targeted miR-221 and negatively regulated miR-221 expression. More importantly, miR-221 mediated the protective effect of NEAT1 knockdown, as evidenced by the restoration of cell viability, cell apoptosis, oxidative stress and neuroinflammation in MPP-induced SH-SY5Y cells. In conclusion, our study suggested that NEAT1 silencing alleviated MPP-induced neuronal damage by sponging miR-221 in SH-SY5Y cells, highlighting the role of NEAT1 as a potential molecular target for PD therapy.
长链非编码RNA（lncRNAs）最近因其参与包括帕金森病（PD）在内的多种人类神经退行性疾病而受到越来越多的关注。本研究的目的是探讨NEAT1在PD中的功能作用及潜在机制。采用qRT-PCR评估NEAT1和miR-221的表达，通过蛋白质免疫印迹法检测Bcl-2和Bax的表达水平。分别采用CCK-8法和流式细胞术测定细胞活力和凋亡情况。分别通过ELISA法和qRT-PCR评估氧化应激和神经炎症的变化。通过双荧光素酶报告基因检测和RNA免疫沉淀检测验证NEAT1与miR-221之间的靶向相互作用。我们的数据支持，MPP处理以剂量和时间依赖性方式提高SH-SY5Y细胞中NEAT1的表达，而NEAT1沉默可缓解MPP诱导的SH-SY5Y细胞活力抑制和细胞凋亡增强。此外，NEAT1沉默可减轻MPP诱导的SH-SY5Y细胞氧化应激和神经炎症的促进作用。NEAT1直接靶向miR-221并负向调节miR-221的表达。更重要的是，miR-221介导了NEAT1敲低的保护作用，MPP诱导的SH-SY5Y细胞中细胞活力、细胞凋亡、氧化应激和神经炎症的恢复证明了这一点。总之，我们的研究表明，NEAT1沉默通过在SH-SY5Y细胞中吸附miR-221减轻了MPP诱导的神经元损伤，突出了NEAT1作为PD治疗潜在分子靶点的作用。
