IL-22对类风湿关节炎成纤维样滑膜细胞凋亡的影响

Effects of IL-22 on Apoptosis of Rheumatoid Arthritis Fibroblast-like Synoviocytes

【目的】探讨IL-22对类风湿关节炎(RA)成纤维样滑膜细胞(FLS)凋亡的影响。【方法】收集行关节置换术或关节镜下滑膜切除术患者的滑膜组织,组织块法培养FLS。利用荧光定量RT-PCR(q RT-PCR)检测Bcl-2 m RNA和Bcl-XL m RNA、Western Blotting检测Bcl-2蛋白在RA、骨关节炎(OA)和创伤(Trauma)患者中FLS的表达水平。将RA-FLS以一氧化氮(NO)供体硝普钠(SNP)和/或重组人源性IL-22(rh IL-22)等处理24 h,流式细胞术分析检测凋亡。rh IL-22以不同的浓度(0、1、10、100 ng/m L)作用于RA-FLS和OA-FLS 24 h,Western Blotting检测Bcl-2蛋白表达水平的变化。【结果】RA-FLS中Bcl-2 m RNA(P<0.05)、Bcl-XL m RNA(P<0.01)和Bcl-2蛋白(P<0.01)的表达水平明显高于与OA-FLS组和Trauma-FLS组。流式细胞术结果显示:与对照组相比,rh IL-22组细胞凋亡率未见明显变化(P>0.05),SNP组RA-FLS细胞凋亡率明显升高(P<0.01);与SNP组相比,SNP+rh IL-22组细胞凋亡率明显下降(P<0.01)。rh IL-22可明显上调RA-FLS和OA-FLS中Bcl-2的蛋白表达水平(P<0.05),但RA-FLS对rh IL-22的作用更敏感。【结论】IL-22可抑制由SNP诱导的RA-FLS细胞凋亡,其机制可能部分与上调RA-FLS中Bcl-2的表达有关。

[ Objective ] To investigate the effect of interleukin （IL）-22 on the apoptosis of rheumatoid arthritis fibroblast-like synoviocytes （RA-FLS）. [Methods] Synovium tissues collected and isolated from patients who had joint replacement surgery or arthroscopy, which were cultured by tissue culture method. Bcl-2 mRNA and Bcl-XL mRNA expression in FLS （RA, Osteoarthritis, Trauma） were determined by qRT-PCR.Bcl-2 protein expression in FLS were detected by Western blotting. RA-FLS were treated with nitric oxide （NO） donor sodium nitroprusside （SNP） in the present or absence of rhIL-22, RA-FLS apoptosis was analyzed by flow cytometry. Effects of different concentrations of rhIL-22（0, 1, 10, 100 ng/mL） on the expression of Bcl-2 protein level in RA- FLS and OA-FLS were detected by Western Blotting. [Results] The Bcl-2 mRNA （P 〈 0.05）, Bel-XL mRNA （P 〈 0.01） and Bcl-2 protein （P〈0.01） were increased in RA-FLS compared with OA-FLS and Trauma-FLS. Flow cytometry assay showed rhIL-22 did not affect RA-FLS apoptosis compared with control group （P 〉 0.05）, SNP induced RA-FLS apoptosis obviously （P 〈 0.01 ）, rhIL-22 prevented RA-FLS apoptosis in the presence of SNP （P 〈 0.01 ）. rhIL-22 could up-regulate the expression of Bcl-2 in both RA-FLS and OA-FLS （P 〈 0.05）, but RA-FLS were more sensitive to rhlL-22. [Conclusion] IL-22 can protect RA-FLS from SNP-indueed apoptosis, at least in partly by up-regulating the expression of Bcl-2.