三氧化二砷对兔卵蛋白诱导性关节炎的治疗作用的实验研究

Arsenious acid experimental study of the therapeutic effect of the rabbit ovalbumin induced rheumatoid arthritis

目的探讨三氧化二砷对日本大耳白兔卵蛋白诱导性关节炎模型外周血中抑炎因子(IL-10)、促炎因子(TNF-α)、CD4+、CD8+T细胞含量及CD4+/CD8+比值的影响。方法将48只日本大耳白兔随机分为模型组、正常对照组、三氧化二砷低剂量组(1.0 mg·kg-1·d-1)、三氧化二砷中剂量组(2.0 mg·kg-1·d-1)、三氧化二砷高剂量组(4.0 mg·kg-1·d-1)、激素组醋酸泼尼松龙(10 mg·d-1),每组8只。成功建立卵蛋白诱导关节炎兔模型后,分别给药2周,采用酶联免疫吸附法(ELISA)检测兔外周血中的IL-10、TNF-α,流式细胞仪检测CD4+、CD8+T细胞的含量。结果卵蛋白诱导法可以成功诱导兔的关节炎,模型组外周血中的IL-10含量较正常对照组降低(P<0.01),模型组TNF-α较正常对照组上升(P<0.05),三氧化二砷各剂量组兔的关节炎症状不同程度改善,IL-10含量较模型组升高(P<0.05),而TNF-α较模型组降低(P<0.05);模型组外周血CD4+及CD4+/CD8+比值较正常对照组显著升高(P<0.01),三氧化二砷治疗组比值均比正常组出现不同程度的下降。结论三氧化二砷可能通过抑制CD4+T细胞的异常增生,减轻细胞免疫应答、调节卵蛋白关节炎兔外周血T细胞各亚群的紊乱缓解RA的病程发展,该作用可能是三氧化二砷治疗类风湿关节炎的重要机理之一。

Objectlve To investigate the effects of arsenious acid on the levels of anti-inflammatory cytokines （IL-10）, proinflammatory cytokines （TNF-et） , CD4 ＋ , CD8 ＋ T ceils and CD4 ＋/ CD8 ＋ ratio in ovalbumin induced rabbit rheumatoid arthritis model. Methods Forty-eight rabbits were randomly assigned into six groups ： positive model group, negative control group, low-dose of Arsenious acid group （1.0 mg . kg-1 . d-1） ,middle-dose of arsenious acid group（2.0 mg . kg-1 d-1） ,high-dose of arsenious acid group（4.0 mg .kg-1 .d- 1 ）and prednisolone group（ 10 mg .d-1 ）. After successful establishment of rabbit model of ovalbumin-induced arthritis, under two weeks of treatment, the concentration of IL-10 and TNF-ain in the, rabbits peripheral blood were detected by enzyme-linked immunosorbent assay（ELISA） ,and the flow cytometry was used to detect the concentration of CD4 ,CD8 ＋ T cells in the rabbits peripheral blood. Results Ovalbumin-induced method can successfully induced rabbit rheumatoid arthritis, the concentration of anti-inflammatory cyto- kines（IL-10） in peripheral blood of the model group was lower than that in control group, and TNF-a was higher than that in control group. In arsenious acid treatment group rabbits arthritis symptoms were improved;and the concentration of IL-10 was elevated compared with the model group （ P 〈 0.05 ）, the TNF-a was sigfiificantly decreased compared with the model group （ P 〈 0.05 ）. The peripheral blood CD4 T cell,CD4 ＋/CD8 T ceils ratios of the model group were significantly higher than those in control group（ P 〈 0.01 ）, and showed varying degrees of decline in arsenious acid treatment groups. Conclusion Arsenic trioxide may reduce the cellular immune response by inhibiting the abnormal proliferation of CD4 T cells, and adjust disorders of T cells in peripheral blood of the ovalbumin arthritis rabbit, ease the progression of RA. The role may be an important mechanism of arsenic trioxide treatment of rheumatoid arthritis.