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
出处
期刊:PubMed
日期:2023-10-01
卷期号:39 (10): 904-909
摘要
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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