IL-27诱导人成纤维样滑膜细胞产生IL-6的机制研究

发布时间：2018-04-14 14:33

本文选题：IL-27 + 类风湿关节炎　；参考：《重庆医科大学》2017年硕士论文

【摘要】：目的:类风湿关节炎是一种以关节软骨损伤和关节滑膜炎症为病理特征的炎症性自身免疫性疾病。成纤维样滑膜细胞是关节滑膜里的一种间充质细胞,在类风湿关节炎的免疫发病机制中发挥着重要的作用。活化的成纤维样滑膜细胞可以产生大量的可溶性的和结合于细胞表面的炎症介质分子,这些炎症介质能导致炎症加剧和组织的破坏。但是,调节成纤维样滑膜细胞释放各种炎症介质的分子机制目前尚未阐述清楚。本研究拟探讨白介素27(Interleukin-27,IL-27)对人成纤维样滑膜细胞(Human fibroblast-like synoviocyte,HFLS)的体外活化效应及细胞内信号传导途径。方法:IL-27刺激人成纤维样滑膜细胞48小时后,ELISA检测细胞培养上清液中IL-6水平的变化;用信号通路抑制剂处理FLS 1小时后,再用IL-27刺激FLS 48小时,ELISA检测细胞上清液中IL-6的水平变化;IL-27刺激FLS不同时间点,western blot检测细胞内信号通路蛋白的磷酸化水平。结果:IL-27能刺激正常关节来源的人FLS和RA患者关节来源的人FLS产生IL-6水平显著升高,且具有时间和剂量依赖性。IL-27能刺激人FLS细胞内信号通路蛋白JAK-2和JNK的磷酸化水平升高。JAK抑制剂AG490和JNK抑制剂SP600125可明显抑制IL-27诱导FLS产生IL-6。结论:IL-27通过激活人FLS细胞内JAK-2和JNK信号通路,从而上调FLS分泌产生IL-6水平,由此说明细胞因子IL-27在类风湿关节炎的发病机制中扮演着重要的促炎症作用。
[Abstract]:Objective: rheumatoid arthritis is an inflammatory autoimmune disease characterized by articular cartilage injury and synovitis.Fibroblast is a kind of mesenchymal cells in synovium, which plays an important role in the immune pathogenesis of rheumatoid arthritis.Activated fibroblast synovial cells can produce a large number of soluble and bound to the surface of the cell surface of inflammatory molecules, these inflammatory mediators can lead to increased inflammation and tissue destruction.However, the molecular mechanisms regulating the release of various inflammatory mediators by fibroid synovial cells have not been clarified.The aim of this study was to investigate the in vitro activation and intracellular signaling pathway of interleukin-27 interleukin-27 (IL-27) on human fibroblast synoviocyte (HFLs) in human fibroblast synoviocytes.Methods the level of IL-6 in the supernatant of cultured human fibroblasts was detected by Elisa after stimulation of human fibroblast synoviocytes by 1: IL-27 for 48 hours, and FLS was treated with signal pathway inhibitor for 1 hour.IL-27 stimulated FLS for 48 hours was used to detect the level of IL-6 in the supernatant. IL-27 stimulated FLS at different time points and western blot was used to detect the phosphorylation level of intracellular signal pathway protein.Results: the level of IL-6 was significantly increased in human FLS from normal joints and FLS in patients with RA.In a time and dose-dependent manner, IL-27 could increase the phosphorylation level of JAK-2 and JNK in human FLS cells. AG490, a JAK inhibitor, and SP600125, an inhibitor of JNK, could significantly inhibit IL-27 induced production of IL-6 in FLS.Conclusion by activating the JAK-2 and JNK signaling pathways in human FLS cells, the cytokine IL-27 up-regulates the secretion of FLS to produce IL-6, which suggests that the cytokine IL-27 plays an important role in promoting inflammation in the pathogenesis of rheumatoid arthritis.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R593.22

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