Intensive Care Unit, Suzhou Ninth People's Hospital, No. 2666, Ludang Road, Taihu New Town, Wujiang District, Suzhou, Jiangsu 215200, China.
Anal Cell Pathol (Amst). 2023 Mar 20;2023:3154223. doi: 10.1155/2023/3154223. eCollection 2023.
Long non-coding RNAs (LncRNAs) OIP5-AS1 and miR-25-3p play important roles in myocardial injury, whereas their roles in lipopolysaccharide (LPS)-induced myocardial injury remain unknown. The purpose of our study was to investigate the functional mechanisms of OIP5-AS1 and miR-25-3p in LPS-induced myocardial injury.
Rats and H9C2 cells were treated with LPS to establish the model of myocardial injury and , respectively. The expression levels of OIP5-AS1 and miR-25-3p were determined by quantitative reverse transcriptase-polymerase chain reaction. Enzyme-linked immunosorbent assay was performed to measure the serum levels of IL-6 and TNF-. The relationship between OIP5-AS1 and miR-25-3p/NOX4 was determined by luciferase reporter assay and/or RNA immunoprecipitation assay. The apoptosis rate was detected by flow cytometry, and cell viability was detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay. Western blot was performed to detect the protein levels of Bax, Bcl-2, caspase3, c-caspase3, NOX4, and p-NF-B p65/NF-B p65.
OIP5-AS1 was up-regulated, and miR-25-3p was down-regulated in myocardial tissues of LPS-induced rats and LPS-treated H9C2 cells. Knockdown of OIP5-AS1 relieved the myocardial injury in LPS-induced rats. Knockdown of OIP5-AS1 also inhibited the inflammation and apoptosis of myocardial cells , which was subsequently confirmed by experiments. In addition, OIP5-AS1 targeted miR-25-3p. MiR-25-3p mimics reversed the effects of OIP5-AS1 overexpression on promoting cell apoptosis and inflammation and on inhibiting cell viability. Besides, miR-25-3p mimics blocked the NOX4/NF-B signalling pathway in LPS-induced H9C2 cells.
Silencing of lncRNA OIP5-AS1 alleviated LPS-induced myocardial injury by regulating miR-25-3p.
长链非编码 RNA（lncRNA）OIP5-AS1 和 miR-25-3p 在心肌损伤中发挥重要作用，但其在脂多糖（LPS）诱导的心肌损伤中的作用尚不清楚。本研究旨在探讨 OIP5-AS1 和 miR-25-3p 在 LPS 诱导的心肌损伤中的功能机制。
分别用 LPS 处理大鼠和 H9C2 细胞，建立心肌损伤模型。采用定量逆转录聚合酶链反应测定 OIP5-AS1 和 miR-25-3p 的表达水平。采用酶联免疫吸附试验测定血清白细胞介素 6（IL-6）和肿瘤坏死因子-α（TNF-α）水平。通过荧光素酶报告基因测定和/或 RNA 免疫沉淀测定确定 OIP5-AS1 与 miR-25-3p/NOX4 的关系。通过流式细胞术检测细胞凋亡率，通过 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐（MTT）比色法检测细胞活力。Western blot 检测 Bax、Bcl-2、caspase3、c-caspase3、NOX4 和 p-NF-B p65/NF-B p65 蛋白水平。
LPS 诱导的大鼠心肌组织和 LPS 处理的 H9C2 细胞中 OIP5-AS1 上调，miR-25-3p 下调。敲低 OIP5-AS1 减轻 LPS 诱导的大鼠心肌损伤。敲低 OIP5-AS1 还抑制了心肌细胞的炎症和凋亡，这随后通过实验得到证实。此外，OIP5-AS1 靶向 miR-25-3p。miR-25-3p 模拟物逆转了 OIP5-AS1 过表达对促进细胞凋亡和炎症以及抑制细胞活力的影响。此外，miR-25-3p 模拟物阻断了 LPS 诱导的 H9C2 细胞中 NOX4/NF-B 信号通路。
沉默长链非编码 RNA OIP5-AS1 通过调节 miR-25-3p 缓解 LPS 诱导的心肌损伤。
