Zhao Mei-Ying, Wang Gui-Qing, Wang Ni-Ni, Yu Qiao-Yan, Liu Rong-Li, Shi Wen-Qian
a Department of Neurology , Zhengzhou Central Hospital Affiliated to Zhengzhou University , Zhengzhou , P.R.China.
b Department of Geriatric Medicine , Zhengzhou Central Hospital Affiliated to Zhengzhou University , Zhengzhou , P.R.China.
Neurol Res. 2019 Jun;41(6):489-497. doi: 10.1080/01616412.2018.1548747. Epub 2019 Apr 23.
Long-non-coding RNAs (lncRNAs) have been involved in central nervous system recently. A number of studies have reported that lncRNA NEAT1 exerts critical roles in neurodegenerative disorder. Beta-site amyloid precursor protein cleaving enzyme 1 (BACE1) has been reported to exert function in the accumulation of amyloid-β (Aβ). Moreover, BACE1 acts as a target of miR-124 in the progression of AD. So far, the biological role and underlying mechanisms of NEAT1 and miR-124 in AD remains elusive.
The relative NEAT1 and miR-124 expression was examined by qRT-PCR in the tissues and cells line of AD. Cell apoptosis was examined by FACS. Luciferase reporter assay was performed to verify that miR-124 is a direct target of NEAT1, and BACE1 is a downstream target of miR-124. qRT-PCR and western blot analysis were also performed to determinate the BACE1 and the phosphorylation of tau protein.
NEAT1 was notably up-regulated and miR-124 was remarkably down-regulated in AD mouse model. Knockdown of NEAT1 or overexpression of miR-124 showed the protective effects on cellular AD model induced by Aβ. Moreover, miR-124 expression could be up- and down-regulated by suppression or overexpression of NEAT1, respectively. In addition, the expression of BACE1 was the potential functional target of miR-124. These findings suggested that NEAT1 might play a vital role in the development of AD by regulating miR-124/BACE1 axis.
The present study showed that NEAT1 worked as a regulating factor to promote the development of AD via modulating miR-124/BACE1 axis, which might be considered as a novel target in AD treatment.
长链非编码RNA（lncRNAs）近来已参与到中枢神经系统中。许多研究报道lncRNA NEAT1在神经退行性疾病中发挥关键作用。据报道，β-位点淀粉样前体蛋白裂解酶1（BACE1）在淀粉样β蛋白（Aβ）的积累中发挥作用。此外，在阿尔茨海默病（AD）进展过程中，BACE1是miR-124的一个靶点。到目前为止，NEAT1和miR-124在AD中的生物学作用及潜在机制仍不清楚。
采用qRT-PCR检测AD组织和细胞系中NEAT1和miR-124的相对表达。通过流式细胞术检测细胞凋亡。进行荧光素酶报告基因检测以验证miR-124是NEAT1的直接靶点，且BACE1是miR-124的下游靶点。还进行了qRT-PCR和蛋白质免疫印迹分析以测定BACE1和tau蛋白的磷酸化水平。
在AD小鼠模型中，NEAT1显著上调，而miR-124显著下调。敲低NEAT1或过表达miR-124对Aβ诱导的细胞AD模型显示出保护作用。此外，分别通过抑制或过表达NEAT1可上调和下调miR-124的表达。另外，BACE1的表达是miR-124的潜在功能靶点。这些发现提示NEAT1可能通过调节miR-124/BACE1轴在AD的发展中发挥重要作用。
本研究表明，NEAT1作为一个调节因子，通过调节miR-124/BACE1轴促进AD的发展，这可能被视为AD治疗中的一个新靶点。
