沉默SMAD 家族成员3( SMAD3) 促进类风湿性关节炎巨噬细胞M2 极化和SMAD7 表达  

Silencing of SMAD family member 3 promotes M2 polarization of macrophagesand the expression of SMAD7 in rheumatoid arthritis

作　　者：费陈晨 谌曦[2] 万磊[2] 范海霞[2] 刘天阳[2] 李明[2] 刘磊[2] 葛瑶[2] 王晴晴 范文杰 周倩 FEI Chenchen;SHEN Xi;WAN Lei;FAN Haixia;LIU Tianyang;LI Ming;LIU Lei;GE Yao;WANG Qingqing;FAN Wenjie;ZHOU Qian(Graduate School,Anhui University of Chinese Medicine,Hefei 230038;Department of Rheumatism Immunity,First Affiliated Hospital,Anhui University of Chinese Medicine,Hefei 230031,China)

机构地区：[1]安徽中医药大学研究生院,安徽合肥230038 [2]安徽中医药大学第一附属医院风湿科,安徽合肥230031

出　　处：《细胞与分子免疫学杂志》2023年第10期904-909,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：安徽省教育厅自然科学研究重点项目(KJ2020A0396)。

摘　　要：目的探讨小干扰RNA(siRNA)沉默SMAD家族成员3(SMAD3)对类风湿性关节炎(RA)巨噬细胞极化及转化生长因子β1(TGF-β1)/SMAD家族信号通路的影响。方法以巨噬细胞与类风湿关节炎成纤维细胞样滑膜细胞(RA-FLS)共培养为细胞模型,TGF-β1刺激巨噬细胞后,将SMAD3特异性siRNA(si-SMAD3)和阴性对照siRNA(si-NC)转染TranswellTM小室共培养的人RA巨噬细胞,实时荧光定量PCR检测SMAD3 mRNA表达,Western blot法分别检测TGF-β1、SMAD3和SMAD7蛋白的表达;ELISA测定细胞培养上清液中TGF-β1、白细胞介素23(IL-23)的含量;CCK-8法检测细胞增殖情况;TranswellTM小室测定细胞的迁移。结果与模型组和si-NC组相比,沉默SMAD3后,RA巨噬细胞中TGF-β1、SMAD3 mRNA和蛋白表达显著下降,IL-23的分泌明显减少,细胞增殖活性及细胞迁移受到抑制,SMAD7高表达。结论敲低SMAD3促进RA巨噬细胞M2极化及SMAD7表达。Objective To investigate the effect of SMAD family member 3(SMAD3)silenced by small interfering RNA(siRNA)on macrophage polarization and transforming growth factorβ1(TGF-β1)/SMAD family signaling pathway in rheumatoid arthritis(RA).Methods RA macrophages co-cultured with rheumatoid arthritis fibroblast-like synoviocytes(RA-FLS)were used as a cell model.TGF-β1 was used to stimulate macrophages,and SMAD3-specific siRNA(si-SMAD3)and negative control siRNA(si-NC)were transfected into human RA macrophages co-cultured in TranswellTM chamber.The expression of SMAD3 mRNA was detected by real-time fluorescence quantitative PCR,and the expression of TGF-β1,SMAD3 and SMAD7 protein was detected by Western blot analysis.The contents of TGF-β1 and IL-23 in cell culture supernatant were determined by ELISA.Cell proliferation was detected by CCK-8 assay.TranswellTM chamber was used to measure cell migration.Results Compared with the model group and the si-NC group,the expression of TGF-β1,SMAD3 mRNA and protein in RA macrophages decreased significantly after silencing SMAD3.In addition,the secretion of IL-23 decreased significantly,and the cell proliferation activity and cell migration were inhibited,with high expression of SMAD7.Conclusion Knockdown of SMAD3 can promote M2 polarization and SMAD7 expression in RA macrophages.

关 键 词：类风湿性关节炎(RA) SMAD家族成员3(SMAD3) SMAD7 巨噬细胞极化 白细胞介素23(IL-23) 转化生长因子β1(TGF-β1) 

分 类 号：R392-33[医药卫生—免疫学] R593.22[医药卫生—基础医学] R364.5