Knockdown of LncRNA H19 Relieves LPS-Induced Damage by Modulating miR-130a in Osteoarthritis
Knockdown of LncRNA H19 Relieves LPS-Induced Damage by Modulating miR-130a in Osteoarthritis
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DOI：
10.3349/ymj.2019.60.4.381
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发表时间：
2019-04-01
影响因子：
2.4
通讯作者：
Zou, Yonggen
中科院分区：
文献类型：
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作者：
Hu, Yi;Li, Sukai;Zou, Yonggen
目的：骨关节炎是一种常见病、多发病，目前尚无确切的治疗方法。长非编码RNA(LncRNAs)已被广泛证实参与了骨性关节炎进展的调控。本研究旨在探讨LncRNAH19在OA中的作用及其机制。材料和方法：采用逆转录定量聚合酶链式反应(RT-qPCR)检测脂多糖(LPS)诱导的C28/I2细胞中H19和microRNA-130a(miR-130a)的表达。CCK-8和流式细胞仪检测细胞存活率和细胞凋亡率。使用Starbase在线软件预测H19与miR-130a之间的可能结合位点。通过荧光素酶报告、RNA下拉和RT-qPCR分析H19与miR-130之间的相互作用。结果：在脂多糖处理的C28/I2细胞中，H19水平显著升高，且呈剂量依赖关系。H19基因敲除可减轻脂多糖诱导的C28/I2细胞的损伤，表现为诱导细胞活力，减少细胞凋亡，减少炎症因子的分泌。此外，H19通过作为miR-130a的分子海绵，负性调控miR-130a的表达。修复miR-130a后，H19对细胞损伤的刺激作用消失。结论：lncRNAH19通过海绵miR-130a加重了脂多糖诱导的C28/I2细胞的损伤，提示了一种新的调控机制和治疗OA的潜在靶点。
Purpose: Osteoarthritis (OA) is a commonly occurring illness without a definitive cure, at present. Long non-coding RNAs (lncRNAs) have been widely confirmed to be involved in the modulation of OA progression. This study aimed to investigate the role and mechanism of lncRNA H19 in OA.Materials and Methods: Abundances of H19 and microRNA-130a (miR-130a) in lipopolysaccharide (LPS)-treated C28/I2 cells were measured by reverse-transcription quantitative PCR (RT-qPCR). CCK-8 and flow cytometry analyses were carried out to assess cell viability and apoptosis. Starbase online software was used to predict the putative binding sites between H19 and miR-130a. Luciferase reporter, RNA pull down, and RT-qPCR were performed to analyze the true interaction between H19 and miR-130a.Results: A notably dose-dependent elevation of H19 levels was observed in LPS-treated C28/I2 cells. Knockdown of H19 ameliorated the injury of LPS-induced C28/I2 cells, reflected by induced viability, decreased apoptosis, and reduced inflammatory factor secretions. Moreover, H19 negatively regulated the expression of miR-130a via acting as a molecular sponge for miR-130a. The stimulatory effects of H19 on cell damage were abolished following the restoration of miR-130a.Conclusion: LncRNA H19 aggravated the injury of LPS-induced C28/I2 cells by sponging miR-130a, hinting a novel regulatory mechanism and a potential therapeutic target for OA.