摘要
Abstract
Objective To explore the effect of TNF-αon the expression of RUNX3 in synovial fibroblasts in patients with rheumatoid arthritis and its mechanism .Methods RASFs were isolated and primary cultured from synovial tissues of patients with rheumatoid arthritis , TNF-αat different concentrations (0, 0.1, 1, 10, 50 ng/mL) was added for 0, 3, 6, 12, and 24 hours respectively , and the relative expression of RUNX 3 mRNA was detected by quantitative real-time PCR. The fibroblasts of the 3rd-8th generations were randomly divided into the RUNX 3 overexpression group and the NC control group, which were infected with the human RUNX 3 adenovirus and the green fluorescent protein ( GFP) control adenovi-rus, respectively, and after 24 h, both groups were treated with 50 ng/mL TNF-αfor 24 h.The real-time fluorescent quan-titative PCR was used to detect the expression of inflammatory factors [interleukin-6 (IL-6), prostaglandin E2 (PGE-2), chemokine CXC-9 ligand, matrix metalloproteinase 2 (MMP-2), and MMP-13] in the two groups.Transwell assay was used to detect cell invasive ability .The fibroblasts of the 3rd-8th generations were randomly divided into the RUNX 3 silen-cing group and NC control group , which were transfected with siRUNX 3 and NC control siRNA by liposome transfection method, respectively, and after 24 h, both groups were treated with 50 ng/mL TNF-αfor 24 h; the real-time fluorescent quantitative PCR was used to detect the expression of IL-6, PGE-2, CXCL-9, MMP-2, and MMP-13.Results The rela-tive expression level of RUNX 3 mRNA in fibroblasts increased gradually with the increase of TNF-αconcentrations and the duration of action , with a dose-and time-dependent manner;the up-regulation of the relative expression of RUNX 3 mRNA was the most significant when TNF-αconcentration was 50 ng/mL and the treatment time was 24 h (all P<0.05).The number of transmembrane cells in the RUNX3 overexpression group and the NC control group was 56 ±4 and 20 ±3, re-spectively (P<0.01).The relative expression of IL-6, PGE-2 and MMP-13 mRNA in the RUNX3 overexpression group was higher than that in the NC control group (P<0.05 or P<0.01).There was no significant difference in the relative ex-pression of CXCL-9 and MMP-2 mRNA between the two groups (P>0.05).The relative expression of IL-6, CXCL-9, MMP-2 and MMP-13 mRNA in the RUNX3 silencing group was lower than that in the control group (P<0.05).There was no significant difference in the relative expression of PGE-2 mRNA between the two groups ( P>0.05).Conclusion TNF-αcan promote the expression of RUNX3 in fibroblasts with a dose-and time-dependent manner, and the increased expressionof RUNX3 can promote the invasion of fibroblasts by regulating the expression of IL -6 and MMP-13.关键词
类风湿性关节炎/滑膜成纤维样细胞/肿瘤坏死因子α/Runt相关转录因子3/炎性因子Key words
rheumatoid arthritis/synovial fibroblast-like cells/tumor necrosis factor-α/Runt-related transcription factor 3/inflammatory cytokine分类
医药卫生
