长链非编码 RNA NEAT1 通过 U2AF2 稳定 Wnt3a，激活 Wnt/β-连环蛋白通路，减轻缺血性脑卒中诱导的损伤。
LncRNA NEAT1 stabilized Wnt3a via U2AF2 and activated Wnt/β-catenin pathway to alleviate ischemia stroke induced injury.
机构信息
Departments of Neurology, Hunan Provincial People's Hospital, The First-Affiliated Hospital of Hunan Normal University, Changsha 410016, Hunan Province, PR China.
Departments of Neurology, Hunan Provincial People's Hospital, The First-Affiliated Hospital of Hunan Normal University, Changsha 410016, Hunan Province, PR China.
出版信息
Brain Res. 2022 Aug 1;1788:147921. doi: 10.1016/j.brainres.2022.147921. Epub 2022 Apr 19.
BACKGROUND
Ischaemic stroke is the leading cause of mortality and disability in the world. LncRNA NEAT1 has been shown to play an important role in ischaemic injury, but the molecular mechanism remains unclear.
METHODS
qRT-PCR was used to determine the expression of lncRNA NEAT1 in OGD/R-induced BV-2 cells. Cell viability was assessed by an MTT assay, and cell apoptosis was assessed by flow cytometry. The expression of related proteins was evaluated by Western blotting and ELISA. The interactions among lncRNA NEAT1, U2AF2 and Wnt3a mRNA was demonstrated by RIP and RNA pulldown assays. XAV-939 was used as an inhibitor of the Wnt/β-catenin pathway.
RESULTS
LncRNA NEAT1 was found to be downregulated in OGD/R-induced BV-2 cells. Overexpression of lncRNA NEAT1 protected BV-2 cells against OGD/R-induced injury. LncRNA NEAT1 enhanced the stability of Wnt3a mRNA via U2AF2. Knockdown of Wnt3a or blockade of the Wnt/β-catenin pathway rescued the effect of lncRNA NEAT1.
CONCLUSIONS
LncRNA NEAT1 protected cells against OGD/R-induced apoptosis and the inflammatory response by activating the Wnt/β-catenin pathway through upregulation of Wnt3a in a U2AF2-dependent manner. LncRNA NEAT1 could be a promising therapeutic candidate for ischaemic stroke treatment in the future.
背景
缺血性中风是世界上导致死亡和残疾的主要原因。LncRNA NEAT1 已被证明在缺血性损伤中发挥重要作用，但分子机制尚不清楚。
方法
qRT-PCR 用于确定 OGD/R 诱导的 BV-2 细胞中 lncRNA NEAT1 的表达。通过 MTT 测定法评估细胞活力，通过流式细胞术评估细胞凋亡。通过 Western blot 和 ELISA 评估相关蛋白的表达。通过 RIP 和 RNA 下拉测定证明 lncRNA NEAT1、U2AF2 和 Wnt3a mRNA 之间的相互作用。XAV-939 用作 Wnt/β-catenin 通路的抑制剂。
结果
发现 lncRNA NEAT1 在 OGD/R 诱导的 BV-2 细胞中下调。lncRNA NEAT1 的过表达可保护 BV-2 细胞免受 OGD/R 诱导的损伤。lncRNA NEAT1 通过 U2AF2 增强 Wnt3a mRNA 的稳定性。Wnt3a 的敲低或阻断 Wnt/β-catenin 通路挽救了 lncRNA NEAT1 的作用。
结论
lncRNA NEAT1 通过上调 U2AF2 依赖性 Wnt3a 激活 Wnt/β-catenin 通路，保护细胞免受 OGD/R 诱导的细胞凋亡和炎症反应。lncRNA NEAT1 可能成为未来缺血性中风治疗的有前途的治疗候选物。
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