丹参多酚酸盐负性调节小鼠骨髓来源树突状细胞成熟及其部分免疫功能  

Salvianolate negatively regulates maturation and immunologic functions of mouse bone marrow-derived dendritic cells

作　　者：宋少华[1] 郭闻渊[1] 傅志仁[1] 傅宏[1] 沈筱芸[2] 刘芳[1] 王正昕[1] 丁国善[1] 

机构地区：[1]第二军医大学长征医院器官移植中心,上海200003 [2]第二军医大学基础部免疫学研究所,上海200433

出　　处：《第二军医大学学报》2009年第12期1333-1337,共5页Academic Journal of Second Military Medical University

基　　金：上海市卫生局青年科研项目(2008Y085)~~

摘　　要：目的:观察丹参多酚酸盐对小鼠骨髓来源树突状细胞(bone marrow-derived dendritic cells,BMDC)成熟及其部分免疫学功能的影响。方法:小鼠BMDC体外培养时加入促成熟剂脂多糖(DCLPS)或同时加入脂多糖及丹参多酚酸盐(DCSV),其中丹参多酚酸盐设3个浓度:低浓度50μg/ml(SVL)、中浓度100μg/ml(SVM)、高浓度200μg/ml(SVH)。流式细胞仪分析各组细胞表面MHC-Ⅱ分子及共刺激分子CD40、CD80、CD86的表达;FITC-Dextran内吞实验分析各组DC抗原吞噬功能;酶联免疫吸附实验(enzyme linked i mmunosorbent assay,ELISA)检测各组DC培养液上清白细胞介素(interleukin,IL)-12 p40水平。[3H]-TdR掺入法检测各组DC体外混合淋巴细胞反应(mixed lymphocyte reactions,MLR)刺激同种反应性T细胞的增殖能力;ELISA法测定MLR上清IL-10、IL-2、干扰素(interferon,IFN)-γ水平。结果:与DCLPS相比,SVL、SVM、SVH细胞表面MHC-Ⅱ分子和共刺激分子表达明显降低,分泌IL-12 p40明显减少,抗原吞噬能力明显升高,刺激同种反应性T细胞增殖的能力显著下降(P<0.05或P<0.01)。与DCLPS相比,SVL、SVM、SVH组MLR上清中IL-2、INF-γ水平明显降低(P<0.05),而IL-10水平差异无统计学意义。结论:丹参多酚酸盐(50～200μg/ml)能够抑制体外培养体系中小鼠BMDC的成熟,并对其部分免疫学功能具有一定的负性调节作用。Objective：To observe the effect of salvianolate on the maturation and immunologic functions of mouse bone marrow-derived dendritic cells（BMDCs）.Methods： Mouse BMDCs were cultured with GM-CSF and IL-4 for 5 days to obtain immature DCs （imDCs）,then the imDCs were cultured in the presence of LPS（1 μg/ml） or LPS（1 μg/ml） plus salvianolate（50,100 and 200 μg/ml） for 48 h to obtain DCLPS,SVL,SVM and SVH.The immunophenotypes and antigen phagocytosis capabilities of imDCs,DCLPS,SVL,SVM and SVH were analyzed by flow cytometry and FITC-Dextran intake.The IL-12 p40 contents in the supernatants were determined by enzyme linked immunosorbent assay（ELISA）.The stimulatory activities of imDCs,DCLPS,SVL,SVM and SVH on allogeneic T cells in mixed lymphocyte reactions（MLR） were tested by incorporation of 3H-TdR;the contents of IL-10,IL-2 and IFN-γ in the supernatants of MLR were examined by ELISA.Results： Compared with DCLPS,SVL,SVM and SVH had lower expression of MHC-Ⅱ,CD40,CD80 and CD86 molecules,secreted less IL-12 p40 in the supernatant,and exhibited stronger antigen phagocytosis capability;moreover,SVL,SVM and SVH showed significantly weaker activity in stimulating proliferation of allogeneic T cells（P〈0.05,0.01）.Compared with DCLPS,SVL,SVM and SVH groups had significantly decreased IL-2 and INF-γ levels in MLR supernatants（P〈0.05）,but there was no significant difference in the secretion of IL-10.Conclusion： Salvianolate（50-200 μg/ml） can inhibit the maturation of mouse BMDCs in vitro,and thus has a negative effect on immunologic function of BMCDs.

关 键 词：丹参多酚酸盐 树突状细胞 免疫功能 

分 类 号：R967[医药卫生—药理学]