Abstract

Introduction

Rheumatoid arthritis (RA) is an autoimmune disease that affects millions of people. Fibroblast-like synoviocytes (FLSs) located in rheumatoid panni play a pivotal role in the formation of RA. The long noncoding RNA (lncRNA) GAS5 is reportedly downregulated in rheumatoid arthritis. However, its detailed mechanism in RA remains to be explored. This study investigated the roles and related mechanisms of GAS5 in RA.

Methods

The expression levels of GAS5, miR-222-3p, and sirtuin 1 (Sirt1) were evaluated by quantitative PCR (qPCR). Cell proliferation was analyzed by CCK-8 and BrdU assays. Cell apoptosis was assessed by flow cytometry and western blotting. Enzyme-linked immunosorbent assay (ELISA) was utilized to evaluate the levels of TNF-α, IL-1β, and IL-6. The interaction between GAS5 or Sirt1 and miR-222-3p was predicted by starBase and validated by dual-luciferase reporter assay.

Results

GAS5 expression was found to be downregulated in the serum samples of RA patients and in RA-FLSs. GAS5 overexpression or the inhibition of miR-222-3p impeded the activity of RA-FLSs by repressing their proliferation and inflammation and by promoting apoptosis. Mechanistically, GAS5 indirectly regulates Sirt1 expression by binding miR-222-3p. Further experiments confirmed that Sirt1 overexpression restored the anti-RA activity of GAS5 under miR-222-3p mimic.

Conclusions

The miR-222-3p/Sirt1 axis was found to be critical for the function of GAS5 in regulating the proliferation, inflammation, and apoptosis of RA-FLSs. These data indicate GAS5 activation as a potential therapeutic strategy for RA progression.

中文翻译：

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LncRNA GAS5通过调节miR-222-3p/Sirt1信号轴减轻类风湿性关节炎

摘要

介绍

类风湿性关节炎 (RA) 是一种影响数百万人的自身免疫性疾病。位于类风湿性羊奶中的成纤维细胞样滑膜细胞 (FLS) 在 RA 的形成中起关键作用。据报道，长链非编码 RNA (lncRNA) GAS5在类风湿性关节炎中被下调。然而，其在 RA 中的详细机制仍有待探索。本研究调查了GAS5在 RA 中的作用和相关机制。

方法

通过定量 PCR (qPCR) 评估GAS5、miR-222-3p 和 Sirtuin 1 (Sirt1)的表达水平。通过CCK-8和BrdU测定分析细胞增殖。通过流式细胞术和蛋白质印迹评估细胞凋亡。酶联免疫吸附试验 (ELISA) 用于评估 TNF-α、IL-1β 和 IL-6 的水平。GAS5或 Sirt1 与 miR-222-3p之间的相互作用由 starBase 预测，并通过双荧光素酶报告基因测定进行验证。

结果

发现 RA 患者的血清样本和 RA- FLS 中的 GAS5表达下调。GAS5过表达或 miR-222-3p 的抑制通过抑制 RA-FLS 的增殖和炎症以及促进细胞凋亡来阻碍 RA-FLS 的活性。从机制上讲，GAS5通过结合 miR-222-3p 间接调节 Sirt1 的表达。进一步的实验证实，在 miR-222-3p 模拟物下，Sirt1 过表达恢复了GAS5的抗 RA 活性。

结论

发现 miR-222-3p/Sirt1 轴对于GAS5在调节 RA- FLS的增殖、炎症和凋亡中的功能至关重要。这些数据表明GAS5激活是 RA 进展的潜在治疗策略。