THH对胶原诱导型关节炎小鼠Th17细胞及细胞因子的影响

Effects of THH on Th17 cells and related cytokines expression in collagen-induced arthritis mice

目的通过观察昆明山海棠(Tripterygium hypoglaucum hutch,THH)对胶原诱导型关节炎(collagen induced arthritis,CIA)小鼠Th17细胞及相关炎性细胞因子的影响,探讨THH治疗类风湿关节炎的相关免疫学机制。方法 30只健康雄性C57BL/6小鼠随机选取10只作为正常对照组,剩余20只用于制备CIA小鼠模型。将造模成功的小鼠随机分为CIA模型组、THH治疗组,每组9只。通过观测小鼠体质量、关节炎指数评分、关节肿胀程度及HE染色了解THH治疗情况,利用流式细胞术、实时荧光定量PCR(RT-qPCR)及ELISA观察THH对CIA小鼠Th17细胞及IL-17A的影响,并使用GraphPad软件对结果进行分析。结果CIA模型组小鼠体质量与THH治疗组间无统计学差异;关节炎指数评分及关节肿胀程度CIA模型组均高于THH治疗组;HE染色显示THH治疗组关节炎的炎症较模型组减轻,骨损伤减少。流式结果显示,脾脏Th17细胞比例CIA模型组低于正常对照组(P<0.01),THH治疗组高于CIA模型组(P<0.05)。CIA模型组踝关节及血清中IL-17A表达明显高于正常对照组(P<0.01),THH治疗组中的表达量低于CIA模型组(P<0.05)。结论 THH可通过升高CIA小鼠外周免疫器官Th17细胞比例、降低关节组织及血清IL-17A表达水平,从而减轻关节炎的炎症反应,减少关节骨损伤。

To investigate the immunologic mechanism of Tripterygium hypoglaucum hutch(THH)in treating rheumatoid arthritis,we observed the effect of THH on Th17 cells and cytokines in collagen-induced arthritis(CIA)mice.Ten of thirty healthy male C57 BL/6 mice were randomly selected as normal control group,and the remaining20 mice were used for preparing CIA mouse model,which were subdivided into CIA control group and THH treatment group.The body weight,score of arthritis index,joint swelling degree and HE staining were observed to understand the treatment status of THH;the effect of THH on Th17 cells in spleen and IL-17 A in ankle joint and serum were detected by flow cytometry,real-time quantitative PCR(RT-qPCR)and ELISA,respectively.GraphPad software was used to analyze the results.Data showed that there was no significant difference in body weight between the CIA control group and the THH treatment group.The score of arthritis index and the degree of joint swelling in the CIA control group were higher than those in the THH treatment group.HE staining showed that the inflammationand the bone damage in the THH treatment group was less than those in the CIA control group.The proportion of Th17 cells in spleen of CIA control group was lower than thsoe in normal control group(P<0.01)and THH treatment group(P<0.05).The mRNA and protein expression levels of IL-17 A in the ankle jointsand serum of CIA control group were significantly higher than that in normal control group(P<0.01)and THH treatment(P<0.05).In conclusion,THH can relieve the inflammatory reaction of arthritis and reduce joint bone damage by increasing proportion of Th17 cell in peripheral immune organ and decreasing the expression of IL-17 A in joint tissue and serum.