丹柴合剂对树突状细胞的诱导分化作用

The differential effects of traditional Chinese medicine Danchaiheji on dendritic cells

目的研究中药复方制剂丹柴合剂对树突状细胞（DCs）的诱导分化作用,并探讨其机制。方法制备丹柴合剂的大鼠含药血清。分离人外周血单个核细胞,经磁珠筛选出CD14＋单核细胞,培养5~7 d获得未成熟树突状细胞（im DCs）,分为空白血清组与含药血清组,空白血清组分别加入含或不含脂多糖（LPS）的大鼠空白血清,含药血清组分别加入含或不含LPS的大鼠含药血清。用流式细胞术检测DCs表面分子CD86、CD11b和人白细胞抗原（HLA）-DR的表达;用酶联免疫吸附试验（ELISA）法检测DCs分泌白细胞介素（IL）-10的水平;用流式细胞术检测DCs对T细胞增殖能力的影响;用实时定量PCR检测吲哚胺2,3双加氧酶（IDO）基因的表达。结果经丹柴合剂作用后,DCs高表达CD11b,低表达CD86与HLA-DR,IL-10分泌增加。且该制剂通过促进DCs表达IDO进而抑制DCs介导的T细胞增殖能力。结论丹柴合剂可诱导DCs分化为调节性树突状细胞（DCregs）并发挥免疫调节作用。

Objective To explore the effects of traditional Chinese formula Danchaiheji on the differentiation of regula-tory dendritic cells（DCs） and the underlying mechanism. Methods The rat blood serums with or without the formula Dan-chaiheji were prepared. The peripheral blood mononuclear cells were separated from the peripheral venous blood of healthydonors. CD14＋monocytes were isolated using CD14＋magnetic beads and cultured for 5-7 days to obtain immature dendriticcells（im DCs）. Then the cells was divided into control group and Danchaiheji containing rat serum group. Control group wasdivided into two subgroups（containing LPS and without LPS）. Danchaiheji containing rat serum group was also divided intotwo subgroups（containing LPS and without LPS）. The surface markers CD86, CD11 b and HLA-DR of DCs were detected byflow cytometry. The level of IL-10 was determined by enzyme-linked immunosorbent assays（ELISA）. The proliferation of al-logeneic T-cells was detected by flow cytometry and the expression level of indoleamine 2,3-dioxygenase（IDO） was deter-mined using quantitative real-time PCR. Results DCs treated with the formula Danchaiheji exhibited high CD11 b and lowCD86 and HLA-DR expression levels as well as promoted the secretion of IL-10. In addition, the drug could inhibit the pro-motion of DCs on the proliferation of T cells, which was associated with the up-regulation of IDO expression. Conclusion The traditional Chinese formula Danchaiheji can induce the differentiation of DCs into regulatory DCs and play a role in in-hibitory effect on immune function.