Diagnosis and Treatment Center of Encephalopathy, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou 510120, China; Department of Neurosurgery, Hospital of Guangzhou University Mega Center, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou 510006, China.
Diagnosis and Treatment Center of Encephalopathy, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou 510120, China.
J Stroke Cerebrovasc Dis. 2020 Oct;29(10):105126. doi: 10.1016/j.jstrokecerebrovasdis.2020.105126. Epub 2020 Jul 15.
Long non-coding RNAs (LncRNAs) have been reported to play important roles in the pathogenesis and development of many diseases, including cerebral ischemia and reperfusion (I/R) injury. In this study, we aimed to investigate the role of LncRNA-Potassium Voltage-Gated Channel Subfamily Q Member 1 opposite strand/antisense transcript 1 (KCNQ1OT1) in cerebral I/R induced neuronal injury, and its underlying mechanisms.
Primary mouse cerebral cortical neurons treated with oxygen-glucose deprivation and reoxygenation (OGD/R) in vitro and mice subjected to middle cerebral artery occlusion (MCAO) and reperfusion were used to mimic cerebral I/R injury. Small inference RNA (siRNA) was used to knockdown KCNQ1OT1 or microRNA-153-3p (miR-153-3p). Dual-luciferase assay was performed to detect the interaction between KCNQ1OT1 and miR-153-3p and interaction between miR-153-3p and Fork head box O3a (Foxo3). Flow cytometry analysis was performed to detect neuronal apoptosis. qRT-PCR and Western blotting were performed to detect RNA and protein expressions.
KCNQ1OT1 and Foxo3 expressions were significantly increased in neurons subjected to I/R injury in vitro and in vivo, and miR-153-3p expression were significantly decreased. Knockdown of KCNQ1OT1 or overexpression of miR-153-3p weakened OGD/R-induced neuronal injury and regulated Foxo3 expressions. Dual-luciferase analysis showed that KCNQ1OT1 directly interacted with miR-153-3p and Foxo3 is a direct target of miR-153-3p.
Our results indicate that LncRNA-KCNQ1OT1 promotes OGD/R-induced neuronal injury at least partially through acting as a competing endogenous RNA (ceRNA) for miR-153-3p to regulate Foxo3a expression, suggesting LncRNA-KCNQ1OT1 as a potential therapeutic target for cerebral I/R injury.
长链非编码 RNA（LncRNA）已被报道在许多疾病的发病机制和发展中发挥重要作用，包括脑缺血再灌注（I/R）损伤。在本研究中，我们旨在研究 LncRNA-钾电压门控通道亚家族 Q 成员 1 反义链/反义转录本 1（KCNQ1OT1）在脑 I/R 诱导的神经元损伤中的作用及其潜在机制。
体外采用氧葡萄糖剥夺再灌注（OGD/R）处理原代小鼠皮质神经元和大脑中动脉闭塞（MCAO）再灌注的小鼠模拟脑 I/R 损伤。使用小干扰 RNA（siRNA）敲低 KCNQ1OT1 或 microRNA-153-3p（miR-153-3p）。双荧光素酶报告基因实验检测 KCNQ1OT1 与 miR-153-3p 的相互作用以及 miR-153-3p 与叉头框 O3a（Foxo3）的相互作用。流式细胞术检测神经元凋亡。qRT-PCR 和 Western blot 检测 RNA 和蛋白表达。
体外和体内 I/R 损伤的神经元中 KCNQ1OT1 和 Foxo3 的表达明显增加，而 miR-153-3p 的表达明显降低。敲低 KCNQ1OT1 或过表达 miR-153-3p 可减弱 OGD/R 诱导的神经元损伤并调节 Foxo3 的表达。双荧光素酶分析表明，KCNQ1OT1 与 miR-153-3p 直接相互作用，Foxo3 是 miR-153-3p 的直接靶标。
我们的结果表明，LncRNA-KCNQ1OT1 通过作为 miR-153-3p 的竞争性内源性 RNA（ceRNA）来调节 Foxo3a 表达，从而促进 OGD/R 诱导的神经元损伤，提示 LncRNA-KCNQ1OT1 可能成为脑 I/R 损伤的潜在治疗靶点。
