摘要目的 探讨人脐带间充质干细胞(HUCMSC)在炎性细胞因子刺激及与类风湿关节炎成纤维样滑膜细胞(RA-FLSs)共培养条件下表达可溶性血管内皮生长因子受体1(sFlt1)水平.方法 ①单独培养HUCMSC,或分别以TNF-α、IFN-γ、TNF-α+IFN-γ刺激HUCMSC,24 h、48 h和72 h后收集上清,ELISA检测各组sFlt1水平.②将RA-FLSs与HUCMSC通过Transwell体系共培养72 h,以RA FLSs和HUCMSC单独培养为对照,ELISA检测各组sFlt1水平.2组间比较采用t检验,多组比较采用单因素方差分析.结果①24 h和48 h时间点,TNF-α+IFN-γ组sFlt1浓度较对照组显著升高[24 h (5.4±0.4) ng/ml和(2.8±1.7) ng/ml,t=0.942,P=0.026;48 h(7.2±-0.8)ng/ml和(4.3±1.0) ng/ml,t=4.285,P=0.005].72 h时间点,TNF-α+IFN-γy组sFlt浓度高于对照组(9.1±1.7) ng/ml和(7.0±1.4) ng/ml,t=0.683,P=0.52),但差异无统计学意义.TNF-α组和IFN-γ组sFlt1浓度较对照组差异无统计学意义.②HUCMSC表达sFlt1水平高于RA-FLSs(8.19±3.64) ng/ml和(0.19±0.08) ng/ml,t=7.280,P＜0.01).RA FLSs与HUCMSC共培养后,共培养组sFlt1浓度显著高于HUCMSC组[(17.26±6.92) ng/ml和(8.19±3.64) ng/ml,t=3.985,P=0.000 7].结论HUCMSC在炎性细胞因子TNF-α联合IFN-γ刺激下表达sFlt1升高.RA FLSs可能通过分泌炎症因子促进共培养HUCMSC上调sFlt1表达.

abstractsObjective To investigate the expression of soluble vascular endothelial growth factor receptor 1 (sFlt1) in human umbilical cord-derived mesenchymal stem cells (HUCMSC) under the condition of inflammatory cytokine and co-culture with rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs).Methods ① HUCMSC were cultured alone or stimulated by Tumour necrosis factor (TNF)-αt,interferon (IFN)-γor the combination of TNF-α and IFN-γ.The sFh1 levels of each group were detected by enzyme-linked immunosorbent assay (ELISA) at 24 h,48 h and 72 h respectively.② RA-FLSs and HUCMSC were cocultured in a transwell system with both of them cultured alone as control for 72 hours.ELISA was used to detect sFlt1 levels in each group.T test was used for comparison between two groups,and ANOVA was used to compare multi-group variables.Results ① The sFlt1 levels of TNF-α + IFN-γ group were significantly higher than those of the control group at 24 h and 48 h (24 h (5.4±0.4) ng/ml vs (2.8±1.7) ng/ml,t=0.942,P=0.026;48 h (7.2±0.8) vs (4.3±1.0) ng/ml,t=4.285,P=0.005),while that at 72 h was not significantly different [(9.1±1.7) ng/ml vs (7.0±1.4) ng/ml,t=0.683,P=0.52].And no significant difference in sFlt1 levels between control group and TNF-α group or IFN-γ stimulation group were observed.② Compared with RA-FLSs,the expression of sFlt1 in HUCMSC was significantly higher (8.19±3.64) ng/ml vs (0.19±0.08) ng/ml,t=7.280,P＜0.01].After co-cultured with RA-FLSs and HUCMSC,the sFlt1 concentration of the co-culture group was significantly higher than that of HUCMSC group [(17.26±6.92) ng/ml vs (8.19±3.64) ng/ml,t=3.985,P=0.000 7].Conclusion The sFlt1 expression of HUCMSC is up-regulated by inflammatory cytokine TNF-α combined with IFN-γ.RA FLSs may promote the co-cultured HUCMSC to increase sFlt1 expression by secreting inflammatory factors.