Zhang Yan, Cao Xu-Qing, Ma Xiao-Li, Guo Yin-Yan, Zhang Tao, Wu Li-Li, Yang Ya-Shan, Hao Chun-Fang, Liu Wei-Li, Guo Jiang-Tao
Department of Rheumatology and Immunology, People's Hospital of Ningxia Hui Autonomous Region (The Third Affiliated Hospital of Ningxia Medical University), Yinchuan City, China.
Department of Neurology, People's Hospital of Ningxia Hui Autonomous Region (The Third Affiliated Hospital of Ningxia Medical University), Yinchuan, China.
Kaohsiung J Med Sci. 2025 Aug;41(8):e70010. doi: 10.1002/kjm2.70010. Epub 2025 Jun 23.
This study examined YTHDF2's role in modulating IL-6R signaling to regulate synovial fibroblast inflammation and bone damage in rheumatoid arthritis (RA). Synovial tissues of RA patients were collected. Human fibroblast-like synoviocyte (FLS), MH7A cell line, was induced with TNF-α and transfected. Cell proliferation was assessed using MTT and EdU assays; apoptosis was measured with flow cytometry, and migration and invasion were evaluated through scratch and Transwell assays. Lentiviral vectors designed to overexpress YTHDF2 or IL-6R were created to study their effects in mice with collagen-induced arthritis (CIA). Pathological changes of ankle joints in mice were observed, and TNF-α, IL-1β, and IL-6 contents were determined. MMP3 and MMP9 levels were detected by Western blot, while YTHDF2 and IL-6R were detected by RT-qPCR and Western blot. The binding relationship between YTHDF2 and IL-6R was studied. YTHDF2 in synovial tissues of RA patients was down-regulated. Elevating YTHDF2 inhibited TNF-α-induced MH7A cell proliferation, migration, invasion, and pro-inflammatory factors; Knocking down YTHDF2 showed the opposite effect. Upregulating YTHDF2 improved synovial inflammation and bone damage in CIA mice. IL-6R in synovial tissues of patients was significantly up-regulated and negatively correlated with YTHDF2 expression. YTHDF2 reduced IL-6R mRNA stability in a m6A-dependent manner. Overexpressing IL-6R impaired the anti-proliferating and anti-inflammatory effect of YTHDF2 on TNF-α-induced MH7A cells. In CIA mice, overexpression of IL-6R reversed the benefits on synovial inflammation and bone injury mediated by up-regulating YTHDF2. YTHDF2 inhibits inflammation and bone damage in RA synovial fibroblasts by reducing the mRNA stability of IL-6R.
本研究探讨了YTHDF2在调节白细胞介素-6受体(IL-6R)信号传导以调控类风湿关节炎(RA)滑膜成纤维细胞炎症和骨损伤中的作用。收集了RA患者的滑膜组织。用人肿瘤坏死因子-α(TNF-α)诱导并转染人成纤维样滑膜细胞(FLS)MH7A细胞系。使用MTT和EdU试验评估细胞增殖;通过流式细胞术检测细胞凋亡,并通过划痕试验和Transwell试验评估细胞迁移和侵袭能力。构建了旨在过表达YTHDF2或IL-6R的慢病毒载体,以研究它们在胶原诱导性关节炎(CIA)小鼠中的作用。观察小鼠踝关节的病理变化,并测定TNF-α、白细胞介素-1β(IL-1β)和IL-6的含量。通过蛋白质免疫印迹法检测基质金属蛋白酶3(MMP3)和基质金属蛋白酶9(MMP9)水平,同时通过逆转录定量聚合酶链反应(RT-qPCR)和蛋白质免疫印迹法检测YTHDF2和IL-6R。研究了YTHDF2与IL-6R之间的结合关系。RA患者滑膜组织中的YTHDF2表达下调。上调YTHDF2可抑制TNF-α诱导的MH7A细胞增殖、迁移、侵袭及促炎因子的产生;敲低YTHDF2则表现出相反的效果。上调YTHDF2可改善CIA小鼠的滑膜炎和骨损伤。患者滑膜组织中的IL-6R显著上调,且与YTHDF2表达呈负相关。YTHDF2以m6A依赖的方式降低IL-6R mRNA的稳定性。过表达IL-6R可削弱YTHDF2对TNF-α诱导的MH7A细胞的抗增殖和抗炎作用。在CIA小鼠中,过表达IL-6R可逆转上调YTHDF2介导的对滑膜炎和骨损伤的有益作用。YTHDF2通过降低IL-6R的mRNA稳定性来抑制RA滑膜成纤维细胞中的炎症和骨损伤。