Guan Yue, Li Xin, Yang Hemin, Xu Siyu, Shi Lidong, Liu Yangyang, Kong Lingdan, Qin Ying
Department of Rheumatology and Immunology, The Third Affiliated Hospital of Qiqihar Medical University, Qiqihar, China.
Central Laboratory, The Third Affiliated Hospital of Qiqihar Medical University, Qiqihar, China.
Heliyon. 2024 Aug 5;10(15):e35589. doi: 10.1016/j.heliyon.2024.e35589. eCollection 2024 Aug 15.
To explore the mechanisms of IRF9 in the progression of rheumatoid arthritis(RA), and the effects of IRF9 on M1/M2 polarization.
RA dataset (GSE55457) was downloaded from GEO. Correlation analysis between IRF9 and its downstream target protein PSMA5 was performed using bioinformatics analysis. The M1/M2 cell ratio of peripheral blood mononuclear cells which from 20 healthy specimen and 40 RA patients was determined. The expression of IRF9 and PSMA5 was detected using qPCR and Western blot. Then, knockdown IRF9 in RAW264.7 cell line (sh-IRF9 RAW264.7) was constructed. The effect of sh-IRF9 RAW264.7 on RA was explored by constructing a CIA mouse model.
IRF9 is upregulated in RA and is of good early screening effect. The results of pathway analysis showed that IRF9 targets and regulates the PSMA5 signaling pathway. IRF9 and PSMA5 were significantly elevated in RA patients, M1/M2 ratio was also increased. The effects of IRF9 on RAW264.7 macrophages were deeply explored in vitro, revealing that knockdown of IRF9 suppressed PSMA5, M1/M2 ratio and the secretion of pro-inflammatory factor in RAW264.7. In mouse in vivo experiments, sh-IRF9 RAW264.7 cells were found to modulate RA by downregulating PSMA5, modulating the M1/M2 ratio through enhancing the anti-inflammatory factor, and suppressing the pro-inflammatory factor.
IRF9 promoted the progression of RA via regulating macrophage polarization through PSMA5.
探讨干扰素调节因子9（IRF9）在类风湿关节炎（RA）进展中的作用机制，以及IRF9对M1/M2极化的影响。
从基因表达综合数据库（GEO）下载RA数据集（GSE55457）。采用生物信息学分析方法对IRF9与其下游靶蛋白蛋白酶体亚基α5（PSMA5）进行相关性分析。测定20例健康标本和40例RA患者外周血单个核细胞的M1/M2细胞比例。采用实时定量聚合酶链反应（qPCR）和蛋白质免疫印迹法检测IRF9和PSMA5的表达。然后，构建RAW264.7细胞系中IRF9基因敲低细胞株（sh-IRF9 RAW264.7）。通过构建胶原诱导性关节炎（CIA）小鼠模型，探讨sh-IRF9 RAW264.7对RA的影响。
IRF9在RA中表达上调，具有良好的早期筛查作用。通路分析结果显示，IRF9靶向并调节PSMA5信号通路。RA患者中IRF9和PSMA5显著升高，M1/M2比例也升高。在体外深入研究了IRF9对RAW264.7巨噬细胞的影响，结果显示敲低IRF9可抑制RAW264.7中的PSMA5、M1/M2比例及促炎因子分泌。在小鼠体内实验中，发现sh-IRF9 RAW264.7细胞可通过下调PSMA5、增强抗炎因子调节M1/M2比例及抑制促炎因子来调节RA。
IRF9通过PSMA5调节巨噬细胞极化促进RA进展。
