| 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50 μg/ml(SVL)、中浓度100 μg/ml(SVM)、高浓度200 μg/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 immunosorbent 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(50~200 μg/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。 |
| 关键词: 丹参多酚酸盐 树突状细胞 免疫功能 |
| DOI：10.3724/SP.J.1008.2009.01333 |
| 投稿时间：2009-07-08修订日期：2009-11-28 |
| 基金项目:上海市卫生局青年科研项目（2008Y085）. |
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| Salvianolate negatively regulates maturation and immunologic functions of mouse bone marrow-derived dendritic cells |
| SONG Shao-hua1△,GUO Wen-yuan1△,FU Zhi-ren1,FU Hong1*,SHEN Xiao-yun2,LIU Fang1,WANG Zheng-xin1,DING Guo-shan1 |
| (1.Organ Transplantation Center,Changzheng Hospital,Second Military Medical University,Shanghai 200003,China;2.Institute of Immunology,College of Basic Medical Sciences,Second Military Medical University,Shanghai 200433) |
| Abstract: |
| 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. |
| Key words: salvianolate dendritic cells immunologic function |
