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目的探讨条件免疫反应治疗胶原性关节炎(CIA)免疫学机制。方法建立大鼠CIA模型。将模型大鼠分为5组:条件免疫反应(CIR)组:以樟脑气味为条件刺激;甲氨蝶呤(MTX)+泼尼松(Pred)为非条件刺激,两者结合7次(7d)后可建立条件免疫反应,然后每日再现条件刺激,每周条件刺激与非条件刺激结合1次,共4周。MTX+Pred组:MTX+Pred治疗4周。MTX+Pred减量组:MTX+Pred治疗,7d内每天1次,7d后每周1次,共4周。单纯闻樟脑气味组:单纯闻樟脑气味4周。空白对照组:安慰剂治疗4周。用流式细胞技术检测大鼠外周血CD4+T淋巴细胞活化(表达CD71)状态,免疫散射比浊法检测大鼠外周血IgG、IgA、IgM、C3、C4、C反应蛋白(CRP)水平,免疫组织化学检测CD68(巨噬细胞标志)及肿瘤坏死因子(TNF)-α在滑膜组织的表达。结果CIR组与MTX+Pred组大鼠治疗2周后外周血活化的CD4+T淋巴细胞明显低于其他各组(P<0.01);4周后血清IgG、CRP水平降低,滑膜组织中CD68及TNF-α表达均明显低于单纯闻樟脑气味组、MTX+Pred减量组及空白对照组(P均<0.01)。结论以樟脑气味为条件刺激,以泼尼松和甲氨蝶呤联合免疫抑制作用为非条件刺激,通过强化训练,建立的条件免疫抑制反应可以通过调节机体的细胞免疫与体液免疫功能,抑制免疫细胞异常活化,降低自身抗体及炎性细胞因子水平,
Objective To investigate the immune mechanism of conditional immune response in treatment of collagen-induced arthritis (CIA). Methods Rat CIA model was established. The model rats were divided into 5 groups: conditioned immune response (CIR) group: stimulation with camphor odor; methotrexate (MTX) + prednisone as unconditioned stimuli, ) To establish a conditioned immune response, and then reproduce the daily conditions of stimulation, weekly and non-conditional stimuli combined with a stimulus for a total of 4 weeks. MTX + Pred group: MTX + Pred for 4 weeks. MTX + Pred reduction group: MTX + Pred treatment, once daily for 7 days, once a week for 7 days, for 4 weeks. Smell camphor smell group: Smell camphor smell for 4 weeks. Blank control group: placebo for 4 weeks. Flow cytometry was used to detect the activation of CD4 + T lymphocytes in peripheral blood of rats (express CD71), IgG, IgA, IgM, C3, C4, C-reactive protein (CRP) Immunohistochemistry was used to detect the expression of CD68 (macrophage marker) and tumor necrosis factor (TNF) -α in synovial tissue. Results The levels of CD4 + T lymphocytes in peripheral blood of CIR group and MTX + Pred group were significantly lower than those of other groups (P <0.01) after 2 weeks of treatment. Serum IgG and CRP levels were decreased after 4 weeks, while CD68 (P <0.01). The levels of TNF-α and TNF-α were significantly lower than those of simple camphor odor group, MTX + Pred reduction group and blank control group (all P <0.01). CONCLUSIONS: Camphor-induced odor stimulation and prednisone and methotrexate combined immunosuppression are unconditioned stimuli. By intensive training, the established immunosuppressive response can suppress cellular immunity and humoral immune function through modulation of the body’s immune response Abnormal cell activation, reduce autoantibodies and inflammatory cytokines,