基于MAPK通路探讨蒿藤胶囊对Ⅱ型胶原诱导性关节炎大鼠的免疫抑制机制

Immunosuppressive mechanism of Haoteng Capsules in typeⅡcollagen-induced arthritis rats

目的:探讨蒿藤胶囊对Ⅱ型胶原诱导性关节炎模型(CIA)大鼠的免疫抑制机制。方法:采用弗氏完全佐剂与Ⅱ型胶原混合后乳剂诱导大鼠关节炎模型,每周检测大鼠右后足关节肿胀百分率;末次给药后,观察关节滑膜的病理变化,采用流式细胞术检测蒿藤胶囊对外周血中CD4^(+)^(+)T淋巴细胞的影响,酶联法检测蒿藤胶囊对血清中TNF-α、IL-17、IL-6、IL-21及IL-22的影响,PCR法测定蒿藤胶囊对脾脏组织中JNK、p38、ERK mRNA表达的影响,免疫组化测定蒿藤胶囊对脾脏组织中JNK、p38、ERK蛋白表达的影响,Western blot法测定关节滑膜中JNK、ERK蛋白表达。结果:蒿藤胶囊0.193 g·kg^(-1)在给药14 d时可显著缓解CIA大鼠的关节肿胀率,降低病理学评分及血清炎症因子IL-17、IL-6的水平,抑制CD4^(+)T淋巴细胞升高,下调脾脏组织JNK、p38、ERK mRNA及JNK、ERK蛋白的表达并下调关节滑膜中JNK、ERK蛋白表达。结论:蒿藤胶囊治疗CIA大鼠的免疫抑制机制可能与下调脾脏组织与关节滑膜中JNK、ERK表达,降低外周血中CD4^(+)T淋巴细胞的比例与血清炎症因子IL-17、IL-6的水平有关。

OBJECTIVE To investigate the immunosuppressive mechanism of Haoteng Capsules in typeⅡcollagen-induced arthritis rats.METHODS Rheumatoid arthritis(RA)rat model was induced by the mixture of Freund’s complete adjuvant and typeⅡcollagen emulsion.The percentage of right posterior foot swelling was examined weekly,and after the last administration,the pathological changes of the synovium of joints were observed.Flow cytometry was used to detect the effect of Haoteng Capsules on CD4^(+)T cells in peripheral blood,enzyme linking method was used to detect the effect of Haoteng Capsules on the contents of TNF-α,IL-17,IL-6,IL-21 and IL-22 in serum,immunohistochemistry was used to detect the effect of Haoteng Capsules on the expression levels of JNK,p38 and ERK in spleen tissue,and Western blot was used to detect the JNK and ERK protein expression in joint synovium.RESULTS Haoteng capsules at the dose of 0.193 g·kg^(-1)could significantly relieve the joint swelling rate of CIA rats after 14-day administration,decrease pathological score and serum inflammatory cytokines IL-17 and IL-6,inhibit the increase of CD4^(+)T lymphocytes,down-regulate the expression of JNK,p38 and ERK in spleen tissue and down-regulate the expression of JNK and ERK protein in joint synovium.CONCLUSION Haoteng capsules can inhibit the immune response of CIA rats,the mechanism may be related to the down-regulation of JNK,p38 and ERK protein expression in spleen and joint synovium,and decreasing CD4^(+)T lymphocytes in peripheral blood and serum inflammatory cytokines IL-17 and IL-6.