Long Non-coding RNA THRIL Mediates Cell Growth and Inflammatory Response of Fibroblast-Like Synoviocytes by Activating PI3K/AKT Signals in Rheumatoid Arthritis
Long Non-coding RNA THRIL Mediates Cell Growth and Inflammatory Response of Fibroblast-Like Synoviocytes by Activating PI3K/AKT Signals in Rheumatoid Arthritis
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DOI：
10.1007/s10753-020-01189-x
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发表时间：
2020-03-30
期刊：
影响因子：
5.1
通讯作者：
Ding, Chao
中科院分区：
文献类型：
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作者：
Liang, Yongjian;Li, He;Ding, Chao
本研究探讨了肿瘤坏死因子-α（TNF-α）和异质核核糖核蛋白L（hnRNPL）相关的免疫调节性lncRNA浆细胞瘤变异易位1（THRIL）在类风湿关节炎（RA）中的可能作用及其机制。从RA患者中收集血清样品。从滑膜组织中分离原代成纤维细胞样滑膜细胞（FLS），并通过分离不同的载体进行培养用于后续的细胞实验。采用qRT-PCR分析来评估血清中THRIL的水平。采用酶联免疫吸附试验（ELISA）检测炎症细胞因子水平。MTT法检测细胞活力，Annexin V-FITC/PI凋亡检测细胞凋亡。此外，通过蛋白质印迹法检测凋亡蛋白和通路相关蛋白的水平。Pearson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患者血液中THRIL过表达（p < 0.05）。RA血液样本中的THRIL水平与TNF-α水平、DAS 28和ESR正相关（p < 0.001）。TNF-α处理显著抑制细胞活力并增强细胞凋亡。此外，它提高了IL-1 β和MMP-3的水平（p < 0.05），而THRIL的抑制逆转了TNF-α处理的RA-FLS中的这些作用（p < 0.05）。此外，降低的THRIL显著降低TNF-α处理的RA-FLS中p-PI 3 K和p-AKT的表达（p < 0.05）。本研究提示THRIL可通过激活PI 3 K/AKT信号通路调节FLS的细胞生长和炎症反应，在RA的发生发展中发挥重要作用。
The present study explored the possible functions and the underlying mechanism of tumor necrosis factor-alpha (TNF-alpha) 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-alpha levels, DAS 28, and ESR (p < 0.001). TNF-alpha treatment significantly inhibited cell viability and enhanced cell apoptosis. Furthermore, it elevated the levels of IL-1 beta and MMP-3 (p < 0.05), whereas the suppression of THRIL reversed these effects in TNF-alpha-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-alpha-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.