[Silencing of SMAD family member 3 promotes M2 polarization of macrophages and the expression of SMAD7 in rheumatoid arthritis].

SMAD公司 基因敲除 小干扰RNA 基因沉默 转化生长因子 巨噬细胞极化 细胞生长 转染 分子生物学 免疫印迹 化学 细胞 分泌物 下调和上调 细胞培养 癌症研究 巨噬细胞 细胞生物学 生物 细胞凋亡 体外 基因 生物化学 遗传学
作者
Chenchen Fei,Xi Shen,Lei Wan,Haixia Fan,Tianyang Liu,Ming Li,Lei Liu,Yao Ge,Qingqing Wang,Wenjie Fan,Qian Zhou
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期刊:PubMed 卷期号:39 (10): 904-909
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Objective To investigate the effect of SMAD family member 3(SMAD3) silenced by small interfering RNA (siRNA) on macrophage polarization and transforming growth factor β1 (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.

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