天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (3): 318-321.doi: 10.11958/20150255

• 实验研究 • 上一篇    下一篇

丹柴合剂对树突状细胞的诱导分化作用

李颖曦1,陈丹2,王小东3,景亚青4,李克秋4,李光3   

  1. 1 天津医科大学基础医学院生物学教研室（邮编 300070）， 2药理学教研室
  • 收稿日期:2015-10-23 修回日期:2015-10-30 出版日期:2016-03-15 发布日期:2016-03-15
  • 通讯作者: 李光 E-mail:tmuliying@163.com
  • 作者简介:李颖曦（1990）， 女， 硕士在读， 主要从事免疫耐受方面研究
  • 基金资助:
    国家高技术研究发展计划（863 计划）资助项目（2012AA021003）；国家自然科学基金资助项目（21177091）

The differential effects of traditional Chinese medicine Danchaiheji on dendritic cells

LI Yingxi1, CHEN Dan2, WANG Xiaodong2, JING Yaqing1, LI Keqiu1, LI Guang1△   

  1. 1 Department of Biology, Basic Medical College, Tianjin Medical University, Tianjin 300070, China; 2 Department of Pharmacology, Basic Medical College
  • Received:2015-10-23 Revised:2015-10-30 Published:2016-03-15 Online:2016-03-15
  • Contact: LI Guang E-mail:tmuliying@163.com

PDF (PC)

4161

摘要： 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（imDCs），分为空白血清对照组与含药血清组，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HLA-DR的表达，用酶联免疫反应（ELISA）法检测DCs分泌IL-10的情况，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 丹柴合剂可将诱导DCs分化为DCregs并发挥免疫调节作用。

关键词: 树突状细胞, 调节性树突状细胞, 免疫耐受, 免疫调节, 吲哚胺-吡咯 2,3-双加氧酶

Abstract: Objective To explore the effects of traditional Chinese formula Danchaiheji on the differentiation of regulatory dendritic cells (DCregs) and the underlying mechanism. Methods The rats’ serum with or without the treatment of formula Danchaiheji was prepared. The peripheral blood mononuclear cells were separated from the peripheral venous blood of healthy donors. CD14+ monocytes were isolated using CD14+ magnetic beads and cultured for 5–7 days to obtain immature dendritic cells (imDCs). Then the cells was divided into 4 groups, including the group of Danchaiheji containing rats’ serum after treatment of drug in the presence or absence of lipopolysaccharide (LPS) and the control group containing equal column rats’ serum in the presence or absence of LPS. Then the surface markers CD86, CD11b and HLA-DR of DCs were detected by flow cytometry and the levels of IL-10 were determined by enzyme-linked immunosorbent assays (ELISA). The proliferation of allogeneic T-cells was detected by flow cytometry and the expression level of indoleamine 2,3-dioxygenase (IDO) was determined using quantitative real-time PCR. Results DCs treated with the formula Danchaiheji exhibited high CD11b and low CD86 and HLA-DR expression levels as well as promoted the secretion of IL-10. In addition, the drug could inhibit the promotion of DCs on the proliferation of T cells, which is associated with the upregulation of IDO expression. Conclusion The traditional Chinese formula Danchaiheji could program DCs towards a regulatory DCs fate and the inhibitory effect on immune function was found.

Key words: dendritic cells, regulatory dendritic cells, immune tolerance, immunomodulation, IDO