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目的：初步探讨IFITM3对类风湿关节炎（RA）滑膜成纤维细胞功能的影响。方法：构建RA-FLSs体外细胞模型，通过Western blot检测RA-FLSs中IFITM3的蛋白表达差异；将实验分为Control组、脂多糖（LPS）组、LPS+siNC组、LPS+siIFITM3组、LPS+siIFITM3+C16-PAF组。采用Transwell试验和划痕试验分别观察RA-FLSs侵袭和迁移能力的变化；ELISA检测TNF-α、IL-1、IL-6水平；流式细胞术检测RA-FLSs凋亡情况；CCK8检测细胞增殖能力；用实时荧光定量PCR检测IFITM3、Bax、Bcl-2 mRNA的表达；Western blot检测IFITM3、MEK、ERK、p-ERK1/2、MMP9、Bax、Bcl-2在蛋白水平的表达；在LPS+siIFITM3组，使用C16-PAF激活MER/ERK信号通路，用CCK8检测细胞增殖能力。结果：在本研究中，LPS处理的RA-FLSs中IFITM3表达水平升高（P<0.05）。与LPS+siNC组比较，LPS+siIFITM3组RA-FLSs细胞迁移能力、侵袭能力降低，炎症因子IL-1、IL-6表达降低，细胞凋亡率增加（P<0.05），同时细胞增殖活性降低；细胞中Bax表达增多，Bcl-2、p-ERK、MMP9蛋白表达减少（P<0.05）。与LPS+siIFITM3组比较，LPS+si-IFITM3+C16-PAF组RA-FLSs增殖活性升高（P<0.05）。结论：下调IFITM3可抑制RA-FLSs迁移侵袭能力和炎症因子IL-1、IL-6的表达，抑制增殖、促进凋亡；其调控增殖机制可能与MEK/ERK信号激活水平有关。
Abstract：Objective: To investigate the effect of IFITM3 on the function of synovial fibroblasts in rheumatoid arthritis. Methods: In vitro cell models of RA-FLSs were constructed, and the protein expression differences of IFITM3 in RA-FLSs were detected by Western blot. The experiments were divided into the Control group, the LPS group, the LPS+siNC group, the LPS+siIFITM3 group, and the LPS+siIFITM3+C16-PAF group. Transwell test and scratch test were used to observe the changes in the invasion and migration ability of RA-FLSs, respectively. The levels of TNF-α, IL-1 and IL-6 were detected by ELISA. Flow cytometry was used to detect the apoptosis of RA-FLSs. CCK8 was used to detect cell proliferation. The mRNA expressions of IFITM3, Bax and Bcl-2 were detected by real-time PCR. Western blot was used to detect the protein expressions of IFITM3, MEK, ERK, p-ERK1/2, MMP9, Bax and Bcl-2. In the IFITM3 siRNA panel, the MER/ERK signaling pathway was activated with C16-PAF, and cell proliferation was detected with CCK8. Results: In this study, the expression level of IFITM3 in LPS-treated RA-FLSs was increased(P <0.05). Compared to the LPS group and the LPS+siNC group, the migration ability and invasion ability of RA-FLSs cells in the LPS+siIFITM3 group decreased, the apoptosis rate increased, the expression of inflammatory cytokines IL-1, IL-6 decreased, and the cell proliferation activity decreased(P<0.05). The expression of Bax increased and the expression of Bcl-2, p-ERK and MMP9 decreased(P<0.05). Compared to the LPS+siIFITM3 group, the proliferative activity of RA-FLSs in the LPS+si-IFITM3+C16-PAF group was increased(P<0.05). Conclusion: Down-regulation of IFITM3 can inhibit the migration and invasion ability of RA-FLSs, the expression of inflammatory cytokines IL-1 and IL-6, inhibit proliferation and promote apoptosis. The mechanism of regulation of proliferation may be related to the level of MEK/ERK signal activation.
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基本信息:
DOI：10.13210/j.cnki.jhmu.20250219.002
中图分类号:R593.22
引用信息:
[1]赵芳,刘俊,李星一,等.IFITM3对类风湿关节炎滑膜成纤维细胞功能影响的研究[J].海南医科大学学报,2026,32(02):81-88.DOI:10.13210/j.cnki.jhmu.20250219.002.
基金信息:
国家自然科学基金（82460321）~~
2025-02-19
2025-02-19
2025-02-19