补阳还五汤对致脑炎性T细胞的免疫调节作用研究

The immunoregulative effect of Buyang Huanwu decoction on the encephalitogenic T cell

目的:探讨补阳还五汤对实验性自身免疫性脑脊髓炎(EAE)致脑炎性T细胞的免疫调节作用。方法:C57BL/6雌性小鼠,采用髓鞘少突胶质细胞糖蛋白35-55(MOG35-55)诱导建立EAE模型,免疫后第9天分离脾脏T淋巴细胞,并培养。设补阳还五汤组和PBS对照组(对照组)。48 h后流式细胞术检测T细胞亚群的变化,检测CD^4+CD25^+、CD4^+IL-10^+、CD4^+IFN-γ^+的表达。ELISA法测定上清中IL-10、IL-1β、IL-17、TNF-α的水平;Western blot法检测T细胞CCL-20、MIP-1α、IFN-γ的表达。结果:与对照组比较,补阳还五汤组可以减少CD4^+IFN-γ^+T细胞,增加CD4^+CD25^+、CD4^+IL-10^+T细胞;抑制炎性细胞因子IL-1β、IL-17、TNF-α、IFN-γ的释放,同时促进抗炎性细胞因子IL-10的释放,抑制化学趋化因子CCL-20、MIP-1α的表达。结论:补阳还五汤具有转化炎性Th1型为抗炎的Th2型及调节性T细胞,抑制炎性反应,降低T细胞趋化因子的表达,提示补阳还五汤具有治疗EAE的潜能。

Objective：To explore the immunoregulative effect of Buyang Huanwu decoction on the encephalitogenic T cell in experimental autoimmune encephalomyelitis mices. Methods：Female adult C57BL/6 mices were immunized with MOG35-55 to induce chronic EAE. Splenic T cells from mice immunized with MOG35-55 on day 9 post-immunization were obtained and treated with Buyang Huanwu decoction or PBS in vitro for 48 h,which were divided into PBS control group and Buyang Huanwu decoction group. Then cells were isolated and the expression of CD4^＋CD25^＋、CD4^＋IL-10^＋、CD4^＋IFN-γ^＋ was analyzed by flow cytometry. The production of IL-10、IL-1β、IL-17、TNF-α was measured by ELISA. We determined the expressions of CCL-20、MIP-1α、IFN-γ by Western blotting. Results：Buyang Huanwu decoction inhibited the expression of CD4^＋IFN-γ^＋T cells and increasing the expression of CD4^＋CD25^＋、CD4^＋IL-10^＋T cells,accompanied by the inhibition of IL-1β、IL-17、TNF-α、IFN-γ and enhanced the level of IL-10. Buyang Huanwu decoction also can reduced the expression of chemokines. Conclusion：Buyang Huanwu decoction can shift inflammatory Th1 to anti-inflammatory Th2 and Treg,inhibit inflammatory responses and reduce the expression of chemokines. Buyang Huanwu decoction has the potential for the treatment of EAE.