IL-22对类风湿关节炎成纤维样滑膜细胞的作用

Effects of IL-22 on rheumatoid arthritis fibroblast-like synoviocytes

目的:探究白细胞介素(IL)-22对类风湿关节炎(RA)成纤维样滑膜细胞(FLSs)功能的影响及机制。方法:组织块法培养RA-FLSs。将不同浓度(0、1、10、100μg/L)的重组人源性IL-22(rhIL-22)与RA-FLSs共培养24 h、48 h、72 h,CCK-8法检测细胞活力的改变;利用10μg/L的rhIL-22作用于RA-FLSs 24 h,流式细胞术检测细胞周期改变。rhIL-22和/或信号转导和转录因子3(STAT3)特异性抑制剂STA-21以不同浓度作用RA-FLSs 24h,Western blot法检测Bcl-2和p-STAT3蛋白水平的变化。结果:不同浓度的rhIL-22作用于RA-FLSs 24 h、48 h、72h后,RA-FLSs细胞活力明显增高,均显著高于对照组(P<0.05)。rhIL-22刺激RA-FLSs后,S期和G_2/M期细胞明显增多,G_0/G_1期细胞减少。Western blot法检测结果提示rhIL-22可上调RA-FLSs中Bcl-2、p-STAT3的蛋白水平,而STA-21单用或联用rhIL-22均可抑制RA-FLSs中Bcl-2及p-STAT3的表达(P<0.05)。结论:IL-22在RA-FLSs细胞活力和周期调节中起重要作用,且STAT3在IL-22促RA-FLSs细胞Bcl-2表达的过程中起关键作用,提示IL-22可能对RA-FLSs凋亡有一定的影响。

AIM： To determine the effects and mechanisms of interleukin-22（ IL-22） on the fibroblast-like synoviocytes（ FLSs） from rheumatoid arthritis（ RA） patients. METHODS： RA-FLSs were cultured by tissue culture method. RA-FLSs were incubated with different concentrations of IL-22（ 0,1,10,100 μg/L） for 24 h,48 h and 72 h. The cell viability was examined by CCK-8 assay. IL-22 at concentration of 10 μg/L was used to stimulate RA-FLSs for 24 h,and the change of cell cycle distribution was identified by flow cytometry. The effects of IL-22 at concentrations of 0,1,10,100 μg/L and/or STA-21（ a STAT3 inhibitor at concentrations of 0,25,50 μmol/L） on the protein levels of Bcl-2 and pSTAT3 in the RA-FLSs were determined by Western blot. RESULTS： Compared with control group,stimulation of rhIL-22 at different concentrations for 24 h,48 h and 72 h,the cells viabilityof RA-FLSs were obviously increased（ P〈 0. 05）.After co-cultured with 10 μg/L rhIL-22 for 24 h,the percentages of RA-FLSs were obviously increased in the G_2/M ＋ S phase and decreased in the G_0/G_1 phase. At the same time,rhIL-22 increased,but STA-21 decreased the protein levels of Bcl-2 but p-STAT3 in the RA-FLSs obviously（ P〈 0. 05）. Treatment with STAT3 inhibitor STA-21 reversed the effect of IL-22-induced Bcl-2 upregulation in the RA-FLSs（ P〈 0. 01）. CONCLUSION： STAT3 is critical in the process of IL-22-induced Bcl-2 upregulation in RA-FLSs,indicating that IL-22 may play a role in the apoptosis of RA-FLSs.