Abstract

The present study explored the possible functions and the underlying mechanism of tumor necrosis factor-α (TNF-α) and heterogeneous nuclear ribonucleoprotein L (hnRNPL)–related immunoregulatory lncRNA plasmacytoma variant translocation 1 (THRIL) in rheumatoid arthritis (RA). Serum samples were collected from patients with RA. Primary fibroblast-like synoviocytes (FLSs) were separated from synovial tissues and cultured for subsequent cell experiments by transfecting different vectors. The qRT-PCR analysis was employed for evaluating the levels of THRIL in the serum. Enzyme-linked immunosorbent assay (ELISA) analysis was employed to detect the levels of inflammatory cytokines. MTT assay and Annexin V-FITC/PI apoptosis assay were used to evaluate the cell viability and apoptosis, respectively. Besides, the levels of the apoptotic proteins and the pathway-related proteins were measured by western blotting. Pearson’s correlation analysis was used to assess the correlation between THRIL and clinical parameters. THRIL was overexpressed in the blood of patients with RA as compared with healthy participants (p < 0.05). The THRIL levels in the RA blood sample were positively associated with TNF-α levels, DAS 28, and ESR (p < 0.001). TNF-α treatment significantly inhibited cell viability and enhanced cell apoptosis. Furthermore, it elevated the levels of IL-1β and MMP-3 (p < 0.05), whereas the suppression of THRIL reversed these effects in TNF-α-treated RA-FLSs (p < 0.05). Moreover, the reduced THRIL remarkably reduced the expression of p-PI3K and p-AKT (p < 0.05) in TNF-α-treated RA-FLSs. The present study revealed that THRIL could regulate cell growth and inflammatory response of FLSs by activating the PI3K/AKT signaling pathway, subsequently playing important roles in promoting the occurrence and development of RA.

中文翻译：

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长的非编码RNA THRIL通过激活类风湿性关节炎中的PI3K / AKT信号介导成纤维细胞样滑膜细胞的生长和炎症反应。

摘要

本研究探讨了类风湿关节炎（RA）中肿瘤坏死因子-α（TNF-α）和异质核糖核糖核蛋白L（hnRNPL）相关的免疫调节lncRNA浆细胞瘤变体1（THRIL）的可能功能及其潜在机制。从RA患者中收集血清样品。从滑膜组织中分离出原代成纤维细胞样滑膜细胞（FLS），并通过转染不同的载体进行培养，以用于随后的细胞实验。使用qRT-PCR分析评估血清中THRIL的水平。酶联免疫吸附测定（ELISA）分析用于检测炎症细胞因子的水平。使用MTT法和膜联蛋白V-FITC / PI凋亡法分别评估细胞活力和凋亡。除了，通过蛋白质印迹法检测凋亡蛋白和通路相关蛋白的水平。皮尔逊相关分析用于评估THRIL与临床参数之间的相关性。与健康参与者相比，RA患者的血液中THRIL过表达（p  ＜0.05）。RA血液样品中的THRIL水平与TNF-α水平，DAS 28和ESR呈正相关（p  <0.001）。TNF-α处理可显着抑制细胞活力并增强细胞凋亡。此外，它提高了IL-1β和MMP-3的水平（p  <0.05），而抑制THRIL则逆转了在TNF-α处理的RA-FLS中的这些作用（p  <0.05）。此外，降低的THRIL显着降低了 TNF-α处理的RA- FLS中p-PI3K和p-AKT的表达（p <0.05）。本研究表明，THRIL可以通过激活PI3K / AKT信号通路来调节FLS的细胞生长和炎症反应，从而在促进RA的发生和发展中起重要作用。