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目的:观察六味地黄丸和金匮肾气丸对实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)小鼠脑组织中肿瘤坏死因子-α(tumor necrosis factor,TNF-α)、转化生长因子-β(transforming growth factor,TGF)-β及白细胞介素(interleukin,IL-17)的调节作用。方法:小鼠皮下多点注射髓鞘少突胶质细胞糖蛋白(myelin oligodendrocyteglycoprotein,MOG)35-55与完全弗氏佐剂(complete Freund’s adjuvant,CFA)的混合抗原,辅以ip百日咳毒素(pertussis toxin,PTX),建立EAE模型。免疫后第12,25,40天取小鼠脑组织,采用免疫组织化学染色法观察各组小鼠脑中TNF-α,TGF-β,IL-17的变化。结果:免疫后第25天,模型组小鼠脑中TNF-α蛋白表达明显高于正常组(P<0.01),而六味地黄丸和金匮肾气丸组则明显低于模型组(P<0.01)。模型组小鼠脑中TGF-β蛋白表达在第40天时明显低于正常组(P<0.01),而各治疗组明显高于模型组(P<0.05或P<0.01),其中六味地黄丸和金匮肾气丸组还显著高于激素组(P<0.05或P<0.01)。模型组小鼠脑中IL-17蛋白表达在第25,40天时均高于正常组(P<0.01),而六味地黄丸和金匮肾气丸组明显低于模型组(P<0.01),并优于激素组(P<0.05)。模型组小鼠脑中TNF-α/TGF-β和IL-17/TGF-β比值在第40天时明显高于正常组(P<0.01),而各治疗组均低于模型组(P<0.01)。结论:六味地黄丸和金匮肾气丸能够调节EAE小鼠中紊乱的TNF-α,TGF-β,IL-17细胞因子,推测可能是其改善EAE病情的机制之一。二方在调节细胞因子的作用方面优于醋酸泼尼松。
Objective: To observe the effects of Liuwei Dihuang Pills and Jingui Shenqi Pill on the expression of tumor necrosis factor-α (TNF-α), transforming growth factor-β β, transforming growth factor (TGF) -β and interleukin (IL-17). Methods: Mice injected subcutaneously with myelin oligodendrocyte glycoprotein (MOG) 35-55 and complete Freund’s adjuvant (CFA) mixed antigen supplemented with ip pertussis toxin toxin, PTX) to establish the EAE model. After 12, 25 and 40 days of immunization, the brain tissues of mice were taken out and the changes of TNF-α, TGF-β and IL-17 in brain were observed by immunohistochemical staining. Results: On the 25th day after immunization, TNF-αprotein expression in the model group was significantly higher than that in the normal group (P <0.01), while that in the Liuweidihuangwan and Jinkuishenqi pills group was significantly lower than that in the model group (P <0.01) ). The expression of TGF-βprotein in the model group was significantly lower than that in the normal group on the 40th day (P <0.01), while the treatment group was significantly higher than the model group (P <0.05 or P <0.01) Kuishen Qi Wan group was also significantly higher than the hormone group (P <0.05 or P <0.01). The expression of IL-17 in brain of model group was higher than that of normal group on the 25th and the 40th day (P <0.01), and was significantly lower in the Liuweidihuangwan and Jinkuishenqi Pills group than in the model group (P <0.01) Superior to hormone group (P <0.05). The ratio of TNF-α / TGF-β and IL-17 / TGF-β in the model group was significantly higher than that in the normal group on the 40th day (P <0.01) ). Conclusion: Liuweidihuang Pill and Jinkuishenqi Pills can regulate the disorganized TNF-α, TGF-β and IL-17 cytokines in EAE mice, suggesting that they may be one of the mechanisms of their improvement in EAE. The two are superior to prednisone acetate in regulating the effects of cytokines.