六味地黄汤通过树突状细胞Notch信号通路调节T细胞分化干预自身免疫性脑脊髓炎

目的 通过观察树突状细胞(dendritic cells,DCs)Notch信号通路对CD4+T细胞分化的影响,探究六味地黄汤干预实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyeli-tis,EAE)的机制.方法 采用主动免疫法诱导2D2小鼠EAE模型,采用磁珠分选收集...

Therapeutic Effect of Liuwei Dihuang Decoction on Autoimmune Encephalomyelitis by Regulating T Cell Differentiation via the Notch Signaling Pathway in Dendritic Cells

Objective To investigate the effect of the Notch signaling pathway in dendritic cells (DCs) on T cell differentiation and the mechanism of Liuwei Dihuang Decoction in intervening against experimental autoimmune encephalomyelitis (EAE). Methods Active immunization was used to induce EAE in 2D2 mice, and magnetic-activated cell sorting was used to collect the CD4+ T cells from the spleen and lymph nodes. Bone marrow-derived dendritic cells (BMDCs) from C57BL/6 (C57) mice were isolated, cultured, and induced to maturity in vitro, and after grouping, intervention, and co-culture with CD4+ T cells, BMDCs were passively injected into C57 mice to observe the onset of the disease. The co-cultured cells of BMDCs and CD4+ T cells were divided into control group, Liuwei Dihuang Decoction group, and gamma-secretase inhibitor (GSI) group. Western blot was used to measure the protein expression levels of the Notch signaling pathway, and flow cytometry was used to analyze the differentiation of CD4+ T cells. Results Compared with the normal group, the model group had significant increases in the protein expression levels of Notch1, Jagged1, and MAML associated with the Notch pathway (P<0.05), and compared with the model group, the Liuwei Dihuang Decoction group had significant reductions in the protein expression levels of Notch1, Jagged1, and MAML (P<0.05). After induction with BMDCs, more CD4+ T cells were differentiated into regulatory T cells, and less CD4+ T cells were differentiated into T helper 17 cells, with an improvement in the symptom score of mice with EAE induced by passive immunization. Conclusion Liuwei Dihuang Decoction can alleviate the disability symptom of EAE by inhibiting the activation of the Notch signaling pathway in DCs and reducing the differentiation of CD4+ T cells.