Department of Orthopaedics, Affiliated Hospital 2 of Nantong University, Nantong University, Nantong 226001, China.
School of Clinical Medicine, Nanjing Medical University, Nanjing 211166, China.
Mediators Inflamm. 2020 Aug 27;2020:4092762. doi: 10.1155/2020/4092762. eCollection 2020.
MicroRNA-155 (miRNA-155) is abundant in fibroblast-like synoviocytes (FLS) in rheumatoid arthritis (RA). Lysine-specific demethylase 1 (LSD1) has been found that it can ameliorate the severity of RA. Tumor necrosis factor-alpha, interleukin-1 beta, and interleukin-6 are key proinflammatory cytokines implicated in the pathogenesis of RA. In our study, we investigated whether miRNA-155 participates in the expression of LSD1 and proinflammatory cytokines in rheumatoid synovial cells. First of all, flow cytometry and cell counting kit-8 analysis were employed to explore the apoptosis and proliferation of FLS, respectively. Subsequently, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was applied to probe into the level of miRNA-155 in FLS when stimulated by miRNA-155 molecules. Moreover, RT-qPCR was used to explore the relative LSD1 miRNA expression in FLS when stimulated by miRNA-155 molecules, and Western blot and immunofluorescence assay were applied to probe into the expression level of LSD1. Finally, enzyme-linked immunosorbent assay was employed to analyze the secreting level of proinflammatory cytokines in FLS when stimulated by miRNA-155 molecules. RA-FLS showed a higher apoptosis rate than normal FLS. The cell proliferation of both HFLS and MH7A cells was promoted by miRNA-155 upregulation. Meanwhile, the expression of LSD1 and proinflammatory cytokines in the FLS of RA was also changed by miRNA-155 regulation. In conclusion, miRNA-155 participates in the expression of LSD1 and proinflammatory cytokines in rheumatoid synovial cells. These findings imply a potential function and interaction of miRNA-155 and LSD1.
微小 RNA-155 (miRNA-155) 在类风湿关节炎 (RA) 的成纤维样滑膜细胞 (FLS) 中含量丰富。赖氨酸特异性去甲基酶 1 (LSD1) 已被发现可以改善 RA 的严重程度。肿瘤坏死因子-α、白细胞介素-1β 和白细胞介素-6 是涉及 RA 发病机制的关键促炎细胞因子。在我们的研究中,我们研究了 miRNA-155 是否参与类风湿滑膜细胞中 LSD1 和促炎细胞因子的表达。首先,采用流式细胞术和细胞计数试剂盒-8 分析分别探讨 FLS 的凋亡和增殖。随后,采用逆转录-定量聚合酶链反应 (RT-qPCR) 探讨 miRNA-155 分子刺激下 FLS 中 miRNA-155 的水平。此外,采用 RT-qPCR 探讨 miRNA-155 分子刺激下 FLS 中 LSD1 的相对 miRNA 表达,采用 Western blot 和免疫荧光法探讨 LSD1 的表达水平。最后,采用酶联免疫吸附试验分析 miRNA-155 分子刺激下 FLS 中促炎细胞因子的分泌水平。RA-FLS 的凋亡率高于正常 FLS。miRNA-155 的上调促进了 HFLS 和 MH7A 细胞的增殖。同时,miRNA-155 调节也改变了 RA FLS 中 LSD1 和促炎细胞因子的表达。综上所述,miRNA-155 参与类风湿滑膜细胞中 LSD1 和促炎细胞因子的表达。这些发现暗示了 miRNA-155 和 LSD1 之间的潜在功能和相互作用。