Wang Zidong, Huang Chuanwang, Zhao Cunju, Zhang Huiling, Zhen Zhen, Xu Duliang
Department of Orthopedic Surgery, Liaocheng People's Hospital, Liaocheng, Shandong 252000, P.R. China.
Department of Endocrinology, Liaocheng People's Hospital, Liaocheng, Shandong 252000, P.R. China.
Exp Ther Med. 2021 Nov;22(5):1244. doi: 10.3892/etm.2021.10679. Epub 2021 Sep 2.
Long non-coding (lnc) RNAs have been associated with osteoarthritis (OA) progression. The aim of the present study was to investigate the regulatory mechanism of lncRNA LINC01385 in OA . The mRNA expression level of LINC01385, microRNA(miR)-140-3p, and Toll-like receptor 4 (TLR4) was detected using reverse transcription-quantitative PCR, while ELISA was used to determine the concentration of different inflammatory factors [tumor necrosis factor-α (TNF-α), IL-6, and prostaglandin E (PGE)]. The viability of human articular chondrocytes (HC-a) was measured using a MTT assay and western blot analysis was performed to quantify the protein expression level of TLR4. The associations between miR-140-3p and LINC01385/TLR4 were confirmed using a dual-luciferase reporter assay. LINC01385 mRNA expression level was increased in OA tissues and IL-1β-induced HC-a. LINC01385 knockdown and miR-140-3p mimics reduced the concentration of inflammatory factors in IL-1β-induced HC-a and promoted cell survival. In addition, it was confirmed that LINC01385 targeted miR-140-3p, while was a target gene of miR-140-3p. Negative correlations between LINC01385 and miR-140-3p, and between miR-140-3p and TLR4 were observed in OA tissues. Low mRNA expression level of miR-140-3p and high protein expression level of TLR4 reversed the inhibitory effect of LINC01385 knockdown on the inflammatory responses of IL-1β-induced HC-a and exhibited a stimulating effect on cell viability. LINC01385 knockdown reduced the progression of OA by modulating the miR-140-3p/TLR4 axis ; thus, LINC01385 may be a therapeutic target for OA.
长链非编码（lnc）RNA与骨关节炎（OA）进展相关。本研究旨在探讨lncRNA LINC01385在OA中的调控机制。采用逆转录定量PCR检测LINC01385、微小RNA（miR）-140-3p和Toll样受体4（TLR4）的mRNA表达水平，同时采用酶联免疫吸附测定法（ELISA）测定不同炎症因子[肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）和前列腺素E（PGE）]的浓度。采用MTT法检测人关节软骨细胞（HC-a）的活力，并进行蛋白质印迹分析以量化TLR4的蛋白表达水平。采用双荧光素酶报告基因测定法证实miR-140-3p与LINC01385/TLR4之间的关联。OA组织和白细胞介素-1β（IL-1β）诱导的HC-a中LINC01385 mRNA表达水平升高。敲低LINC01385和miR-140-3p模拟物可降低IL-1β诱导的HC-a中炎症因子的浓度并促进细胞存活。此外，证实LINC01385靶向miR-140-3p，而TLR4是miR-140-3p的靶基因。在OA组织中观察到LINC01385与miR-140-3p之间以及miR-140-3p与TLR4之间呈负相关。miR-140-3p的低mRNA表达水平和TLR4的高蛋白表达水平逆转了敲低LINC01385对IL-1β诱导的HC-a炎症反应的抑制作用，并对细胞活力表现出刺激作用。敲低LINC01385通过调节miR-140-3p/TLR4轴减少OA的进展；因此，LINC01385可能是OA的治疗靶点。
