目的研究血脑屏障紧密连接蛋白在EAE发病中的作用,观察中药复方益肾达络饮干预实验性自身免疫性脑脊髓炎(EAE)的疗效和作用机制。方法采用荧光定量PCR测定免疫后不同时间点EAE小鼠中枢神经组织中ZO-1 mRNA表达的变化。结果中药复方益肾达络饮可以显著改善EAE模型小鼠神经功能评分。与正常组相比,激素组、中药组在免疫后14、24、40 d ZO-1 mRNA在EAE小鼠大脑中表达均未见显著改变,而脊髓中在免疫后第14日显著降低;在免疫后14、24、40 d模型组小鼠脊髓中ZO-1 mRNA表达均显著降低。结论 ZO-1是实验性自身免疫性脑脊髓炎血脑屏障破坏的一个重要因素;中药复方益肾达络饮能有效改善EAE小鼠神经功能损伤,其干预EAE的作用机制与血脑屏障紧密连接蛋白调节密切相关。
Objective: To elucidate the contribution of ZO-1 in the CNS to the clinical course of EAE mice,and to explore the efficiency and the mechanism of Yishendaluo Decoction to experimental autoimmune encephalomyelitis.Methods: RT-PCR was used to quantify the gene expression levels of ZO-1 in the CNS tissue of mice at different time points of EAE including pre-onset,onset,peak and chronic after immunization.Results: Compared with model group,Yishendaluo Decoction treatment of EAE could improve the neurological function score.Compared with normal group,ZO-1 mRNA levels of the cerebrum in hormone-treated group and Decoction-treated group were not significantly decreased at different time points of EAE including 14,24 and 40 days after immunization,but ZO-1 mRNA levels in the spinal cord in hormone-treated group and Decoction-treated group were significantly decreased only at 14 days.ZO-1 mRNA levels of the spinal cord in hormone-treated group and Decoction-treated group were significantly decreased including 14,24 and 40 days after immunization.Conclusion: ZO-1 plays an important role to the damage of blood-brain barrier in EAE,Treatment with Yishendaluo Decoction can significantly improve the neurological function score.The therapeutic mechanism of Yishendaluo Decoction was to mediate the expression of tight junctions.