Department of Neurology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, China.
Biomed Res Int. 2022 Aug 8;2022:1393177. doi: 10.1155/2022/1393177. eCollection 2022.
Previous studies have reported that lncRNA PVT1 was closely related to ischemic stroke. Here, the role of PVT1 in ischemic stroke and the underlying mechanism were investigated. OGDR-stimulated PC12 cells were used to construct a cell model to mimic ischemic stroke. si-PVT1, miR-214 mimic, inhibitor, or the negative controls were transfected into PC12 cells prior to OGDR treatment. PVT1, miR-214, and Gpx1 expression was measured by qRT-PCR and western blotting assays. Cell proliferation and apoptosis were tested by CCK-8 assay and western blotting. The expression levels of inflammatory factors were determined by ELISA Kit. Results showed that PVT1 was increased significantly in OGDR PC12 cells. PVT1 knockdown significantly enhanced cell viability and attenuated cell apoptosis, ROS generation, and inflammation in OGDR PC12 cells. More importantly, PVT1 or Gpx1 was a target of miR-214. Mechanistically, PVT1 acted as a competing endogenous RNA of miR-214 to regulate the downstream gene Gpx1. In conclusion, PVT1 knockdown attenuated OGDR PC12 cell injury by modulating miR-214/Gpx1 axis. These findings offer a potential novel strategy for ischemic stroke therapy.
先前的研究报道长链非编码 RNA PVT1 与缺血性脑卒中密切相关。本研究旨在探讨 PVT1 在缺血性脑卒中发生发展中的作用及潜在机制。采用氧葡萄糖剥夺（OGDR）刺激 PC12 细胞构建细胞缺血模型。转染 PVT1 小干扰 RNA（si-PVT1）、miR-214 模拟物、抑制剂或阴性对照（NC）至 PC12 细胞，OGDR 处理前进行转染。采用 qRT-PCR 和 Western blot 检测 PVT1、miR-214 和 Gpx1 的表达。CCK-8 法和 Western blot 检测细胞增殖和凋亡。ELISA 试剂盒检测炎症因子的表达水平。结果显示，OGDR 刺激的 PC12 细胞中 PVT1 表达显著增加。与 OGDR 组相比，PVT1 敲低组细胞活力明显增强，细胞凋亡、ROS 生成和炎症反应明显减轻。更为重要的是，PVT1 或 Gpx1 是 miR-214 的靶基因。机制上，PVT1 作为 miR-214 的竞争性内源性 RNA 调节下游基因 Gpx1。综上所述，PVT1 敲低通过调节 miR-214/Gpx1 轴减轻 OGDR PC12 细胞损伤。这些发现为缺血性脑卒中的治疗提供了一种新的潜在策略。
