Dong L I, Zheng Yumin, Gao Lianbo, Luo Xiaoguang
Department of Neurology, The Fourth Affiliated Hospital of China Medical University, Shenyang 110032, China.
Department of Neurology, The First Affiliated Hospital of China Medical University, Shenyang 110000, China.
Acta Biochim Biophys Sin (Shanghai). 2021 Jul 5;53(7):870-882. doi: 10.1093/abbs/gmab055.
Long non-coding RNAs (lncRNAs) play biological roles in brain disorder and neurodegenerative diseases. As the functions of lncRNA NEAT1 in Parkinson's disease (PD) remain unknown, in the present study, we aimed to explore the roles and underlying molecular mechanisms of NEAT1 in PD. A PD mouse model induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and a cell model of SH-SY5Y induced by N-methyl-4-phenylpyridinium (MPP+) were established. The ratio of tyrosine hydroxylase (TH+) cells was determined by immunofluorescence assay, and the behavioral changes in mice were observed using pole tests and rotarod tests. The cellular viability and apoptosis of SH-SY5Y were detected by MTT assay and flow cytometric analysis, respectively, and the number of autophagosomes was subsequently measured by transmission electron microscopy. High-performance liquid chromatography was performed to detect the content of dopamine, and a dual-luciferase reporter assay was used to clarify the target of NEAT1 simultaneously. The results demonstrated that the level of NEAT1 was upregulated in the MPTP-induced PD mice, dopamine neurons, and the SH-SY5Y cells treated with MPP+, whereas the level of miR-374c-5p was downregulated. NEAT1 level was positively correlated with MPP+ in a concentration-dependent manner. NEAT1 inhibition efficiently facilitated cell proliferation but inhibited apoptosis and autophagy in the MPP+-treated SH-SY5Y cells. Additionally, silencing of NEAT1 increased the TH+ rate of neurons and suppressed autophagy greatly in PD mice. As a possible target of NEAT1, miR-374c-5p could impact on the apoptosis and autophagy of the SH-SY5Y cells. NEAT1 inhibition upregulated the expression of miR-374c-5p, enhanced SH-SY5Y cell viability, and repressed autophagy and apoptosis in MPTP-induced PD mice. These findings indicated a potential therapeutic role of NEAT1 in treating PD.
长链非编码RNA（lncRNAs）在脑部疾病和神经退行性疾病中发挥生物学作用。由于lncRNA NEAT1在帕金森病（PD）中的功能尚不清楚，在本研究中，我们旨在探讨NEAT1在PD中的作用及潜在分子机制。建立了由1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）诱导的PD小鼠模型以及由N-甲基-4-苯基吡啶鎓（MPP+）诱导的SH-SY5Y细胞模型。通过免疫荧光测定法测定酪氨酸羟化酶（TH+）细胞的比例，并使用杆式试验和转棒试验观察小鼠的行为变化。分别通过MTT法和流式细胞术分析检测SH-SY5Y细胞的活力和凋亡情况，随后通过透射电子显微镜测量自噬体的数量。采用高效液相色谱法检测多巴胺含量，并同时使用双荧光素酶报告基因检测法阐明NEAT1的靶标。结果表明，在MPTP诱导的PD小鼠、多巴胺神经元以及用MPP+处理的SH-SY5Y细胞中，NEAT1水平上调，而miR-374c-5p水平下调。NEAT1水平与MPP+呈浓度依赖性正相关。抑制NEAT1可有效促进MPP+处理的SH-SY5Y细胞的增殖，但抑制其凋亡和自噬。此外，沉默NEAT1可提高PD小鼠中神经元的TH+比率并极大地抑制自噬。作为NEAT1的可能靶标，miR-374c-5p可影响SH-SY5Y细胞的凋亡和自噬。抑制NEAT1可上调miR-374c-5p的表达，增强SH-SY5Y细胞活力，并抑制MPTP诱导的PD小鼠的自噬和凋亡。这些发现表明NEAT1在治疗PD中具有潜在的治疗作用。
