佐剂性关节炎大鼠IL-23、IL-17检测分析

摘要

Objectives To establish the rheumatoid arthritis (RA) animal models with adjuvant arthritis (AA) rats and to explore the role of IL-23, IL-17 and Thl7 cells in the pathogenesis of AA.Methods Eighteen male SD rats were randomly divided into control and model groups, administered with intradermal injection of 0.1 ml normal saline and 0.1 ml complete Freund's adjuvant, respectively.All rats were killed at day 21 after experiment.The ankle joints were prepared for histopathologic study and the levels of IL-23、 IL-17 in the serum and spleen cell culture supernatant were determined by enzyme-linked immunosorbent assay.Results In model group, an immediate swelling was presented in the injection sites after treating, and a secondary joint swelling appeared in the opposite foot and claws with progressive aggravation, some rats had arthritic nodes in their ears and tails.Pathological changes showed proliferative synovitis in AA rats joints at day 21.Moreover, compared with control group, the levels of IL-23 and IL-17 in model group were obviously higher in the serum (P ＜ 0.01) ; The levels of IL-23 in the spleen cell culture supernatant were obviously higher than that in control group (P ＜ 0.01) , and IL-23 concentrations in the supernatant were significantly higher in the model group after phytohaemagglutinin stimulation (P ＜ 0.01) ; The levels of IL-17 in the spleen cell culture supernatant between two groups were no significant difference, however a pronounced higher level of IL-17 in the spleen cell culture supernatant after PHA stimulation than that in control group (P ＜ 0.01) was found.Conclusions IL-23 and IL-17 might play an important role in the pathogenesis of the AA.Thl7 cells, which was related to IL-23 and produces IL-17, maybe involve in the process of chronic inflammation of AA.%目的 探讨IL-23、IL-17及Th17细胞在类风湿性关节炎发病中的作用,以期对类风湿性关节炎的发病机制和防治对策提供新的认识.方法 将18只SD雄性大鼠随机分为正常对照组及模型组,正常对照组大鼠左后足跖底皮内注射0.1 ml生理盐水,模型组左后足跖底皮内注射0.1 ml完全弗氏佐剂.于造模后15 d确认模型建立成功后,第21天处死大鼠,取踝关节组织,观察病理改变;收集血清并进行脾细胞培养,采用酶联免疫吸附法检测大鼠血清及脾细胞培养上清液中IL-23、IL-17含量.结果 模型组大鼠造模后左足即有肿胀,12 ～ 15 d左右出现右足爪继发性关节肿胀,并进行性加重,部分大鼠耳廓及尾部有关节炎小结出现.造模后 21 d处死大鼠,关节病理切片可见增生性滑膜炎.模型组大鼠血清IL-23及IL-17水平明显升高,与正常对照组比较差异有统计学意义(P ＜ 0.01).模型组脾细胞上清液IL-23水平较对照组显著升高,差异有统计学意义(P ＜ 0.01);经PHA刺激后模型组脾细胞上清液IL-23水平较对照组显著升高,差异有统计学意义(P ＜ 0.01).对照组和模型组脾细胞培养上清液中IL-17差异无统计学意义;PHA刺激后模型组脾细胞培养上清液IL-17水平较对照组显著升高(P ＜ 0.01).结论 IL-23、IL-17是参与佐剂性关节炎发病的重要细胞因子,与IL-23相关、产生IL-17的Th17细胞可能参与了佐剂性关节炎的慢性炎症过程.