Department of Intensive Care Unit (ICU), Xiaogan Central Hospital, Wuhan University of Science and Technology, No. 6 Square Road, South District, Xiaogan, Hubei Province 432000, China.
Department of Anesthesiology, Xiaogan Central Hospital, Wuhan University of Science and Technology, No. 6 Square Road, South District, Xiaogan, Hubei Province 432000, China.
Gene. 2020 May 5;737:144411. doi: 10.1016/j.gene.2020.144411. Epub 2020 Feb 14.
Long non-coding RNAs (lncRNAs) have been identified in cerebral ischemia-reperfusion (I/R) injury nowadays. Herein, we uncovered the function and underlying mechanism of the lncRNA Rian in cerebral I/R injury. The oxygen-glucose deprivation model in N2a cells was offered to mimic cerebral I/R injury in vitro. Trypan blue staining, reactive oxygen species (ROS) production, and caspase-3 activity were used to evaluate cell apoptosis. Then, middle cerebral artery occlusion was conducted to evaluate the function of lncRNA Rian in mice. Real-time PCR and western blotting were performed to determine the expression of lncRNA Rian, miR-144-3p, GATA binding protein 3 (GATA3), caspase-3, Bax, and Bcl-2. The results showed that both Rian and GATA3 were downregulated, and miR-144-3p was upregulated in cerebral I/R injury in vitro and in vivo. Overexpression of Rian could inhibit the cell apoptosis induced by oxygen-glucose deprivation. Furthermore, overexpression of Rian distinctly reduced the infarct size, and it also improved the neurological score. Overexpression of Rian could abolish miR-144-3p-mediated I/R injury in vitro and in vivo. Besides, GATA3 was the target of miR-144-3p and GATA3 could be regulated co-operatively by miR-144-3p and Rian. Consequently, these findings showed that the Rian/miR-144-3p/GATA3 axis is an essential signaling in cerebral I/R injury. The lncRNA Rian may serve as a potential target for novel treatment in patients with ischemic stroke.
长链非编码 RNA（lncRNAs）在脑缺血再灌注（I/R）损伤中已被鉴定出来。本文揭示了 lncRNA Rian 在脑 I/R 损伤中的功能及其潜在机制。在体外采用 N2a 细胞氧葡萄糖剥夺模型模拟脑 I/R 损伤。采用台盼蓝染色、活性氧（ROS）生成和 caspase-3 活性来评估细胞凋亡。然后，进行大脑中动脉闭塞以评估 lncRNA Rian 在小鼠中的功能。采用实时 PCR 和 Western blot 来确定 lncRNA Rian、miR-144-3p、GATA 结合蛋白 3（GATA3）、caspase-3、Bax 和 Bcl-2 的表达。结果表明，Rian 和 GATA3 在体外和体内脑 I/R 损伤中均下调，miR-144-3p 上调。Rian 的过表达可抑制氧葡萄糖剥夺诱导的细胞凋亡。此外，Rian 的过表达明显减少梗死面积，改善神经功能评分。Rian 的过表达可消除 miR-144-3p 介导的体外和体内 I/R 损伤。此外，GATA3 是 miR-144-3p 的靶基因，miR-144-3p 和 Rian 可以协同调节 GATA3。因此，这些发现表明 Rian/miR-144-3p/GATA3 轴是脑 I/R 损伤的重要信号通路。lncRNA Rian 可能成为缺血性脑卒中患者新治疗的潜在靶点。
