Department of Anesthesiology, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, PR China.
Department of Anesthesiology, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, PR China.
Exp Mol Pathol. 2020 Jun;114:104430. doi: 10.1016/j.yexmp.2020.104430. Epub 2020 Mar 30.
Hypoxia/reoxygenation (H/R) injury of cardiomyocytes causes an irreversible damage to heart and largely results in acute myocardial infarction. Study has indicated lncRNA ROR aggravates myocardial ischemia/reperfusion (I/R) injury. Also, lncRNA ROR sponges miR-138 to promote osteogenesis. MiR-138 involves in hypoxic pulmonary vascular remodelling by targeting Mst1. However, the interaction between lncRNA ROR, miR-138 and Mst1 involved in myocardial H/R injury is still unknown.
H9C2 cells were used to establish H/R injury model. The expression levels of lncRNA ROR and miR-138 were modified by transfection with the miR-138 mimics or lncRNA ROR overexpression plasmid. MTT and flow cytometry analysis were performed to detect cell proliferation and apoptosis. Dual luciferase reporter assay was used to determine interaction between lncRNA ROR and miR-138 or miR-138 and Mst1. Expression levels of lncRNA ROR, miR-138, Mst1 and apoptosis-related markers were determined by qRT-PCR or western blotting.
LncRNA ROR was significantly up-regulated, while miR-138 was obviously down-regulated in H/R-induced injury of H9C2 cells. Furthermore, miR-138 overexpression alleviated cardiac cell apoptosis induced by H/R injury. Mst1 was revealed to be a target of miR-138 and negatively regulated by miR-138. Mst1 overexpression reversed the protective effects of miR-138 on H/R injury of H9C2 cells. LncRNA ROR was identified as a sponge for miR-138. MiR-138 could protect H9C2 cells form H/R injury induced by lncRNA ROR overexpression.
Our study provides that lncRNA ROR sponges miR-138 to aggravate H/R-induced myocardial cell injury by upregulating the expression of Mst1.
心肌细胞的缺氧/复氧（H/R）损伤会对心脏造成不可逆的损害，并且在很大程度上导致急性心肌梗死。研究表明长链非编码 RNA ROR 加重心肌缺血/再灌注（I/R）损伤。此外，长链非编码 RNA ROR 通过海绵吸附 miR-138 促进成骨。miR-138 通过靶向 Mst1 参与低氧性肺血管重构。然而，lncRNA ROR、miR-138 和 Mst1 之间的相互作用在心肌 H/R 损伤中的作用尚不清楚。
使用 H9C2 细胞建立 H/R 损伤模型。通过转染 miR-138 模拟物或 lncRNA ROR 过表达质粒来修饰 lncRNA ROR 和 miR-138 的表达水平。采用 MTT 和流式细胞术分析检测细胞增殖和凋亡。双荧光素酶报告基因实验用于确定 lncRNA ROR 与 miR-138 或 miR-138 与 Mst1 之间的相互作用。通过 qRT-PCR 或 Western blot 测定 lncRNA ROR、miR-138、Mst1 和凋亡相关标志物的表达水平。
在 H9C2 细胞的 H/R 损伤中，lncRNA ROR 显著上调，而 miR-138 明显下调。此外，miR-138 过表达减轻了 H/R 损伤引起的心肌细胞凋亡。Mst1 被鉴定为 miR-138 的靶标，并受 miR-138 负调控。Mst1 过表达逆转了 miR-138 对 H9C2 细胞 H/R 损伤的保护作用。lncRNA ROR 被鉴定为 miR-138 的海绵吸附物。miR-138 可以保护 H9C2 细胞免受 lncRNA ROR 过表达引起的 H/R 损伤。
本研究表明，lncRNA ROR 通过上调 Mst1 的表达来海绵吸附 miR-138，从而加重 H/R 诱导的心肌细胞损伤。
