IL-17A对硝普钠诱导的类风湿关节炎成纤维样滑膜细胞凋亡的保护作用及机制研究

Protective effects and mechanisms of IL-17A on SNP-induced apoptosis of fibro-blast-like synoviocytes in patients with rheumatic arthritis

目的：研究IL-17A对一氧化氮（NO）供体硝普钠（SNP）诱导的类风湿关节炎成纤维样滑膜细胞凋亡的保护作用及其信号通路.方法：体外培养类风湿关节炎成纤维样滑膜细胞并以SNP和/或IL-17A等处理;应用MTT法检测细胞存活率;TUNEL染色及流式细胞分析检测凋亡;Western印迹方法检测磷酸化Akt蛋白含量.结果：不同剂量IL-17A（10～100 ng/ml）预处理24小时可浓度依赖性对抗SNP诱导的成纤维样滑膜细胞凋亡,提高细胞存活率;IL-17A上调磷酸化Akt蛋白表达呈浓度与时间依赖性;应用Akt活性抑制剂LY和NF-κB抑制剂PDTC能逆转上述效应.结论：IL-17A抑制了NO供体SNP诱导的类风湿关节炎成纤维样滑膜细胞凋亡,其机制可能部分与激活Akt和NF-κB信号通路相关.

Objective：To study the effects of IL-17A on apoptosis of fibroblast-like synoviocytes （FLS） in patients with rheu- matic arthritis（RA） induced by nitric oxide（NO） donor sodium nitroprusside（SNP） and the possible molecular mechanisms. Meth- ods： Fibroblast-like synoviocytes obtained from patients with active rheumatic arthritis were cultured in vitro and treated with SNP in the present or absence of IL-17A, etc. RA-FLS viability was tested by MTY assay. RA-FLS apoptosis was analysed by flow cytometry and terminal dUTP nick-end labeling （TUNEL）. The levels of phospho-Akt （p-Akt） protein were detected by Western blot. Results： A 24 h IL-17A pretreatment at concentration ranging from 10 - 100 ng/ml increased the viability rate of RA-FLS and prevented RA-FLS apoptosis in a dose-dependent manner in the presence of SNP. IL-17A induced phosphorylation of Akt in a time- and concentration- de- pendent manner. The effect was reversed by treatment with PI3K inhibitor LY（LY） and nuclear factor-KB inhibitor pyrrolidine-dithio- carbamate （PDTC）. Conclusion： IL-17A can protect RA-FLS from SNP-induced apoptosis, at least in partly by activation of Akt and NF-κB signal transduction pathway.