红活麻乙酸乙酯有效部位对小鼠骨髓来源树突状细胞的影响

Effects of the immune-inhibitory active component of traditional Chinese medicine on murine BMDCs

目的观察湖北民族药红活麻乙酸乙酯有效部位对体外培养树突状细胞(DC)功能及表达的影响。方法用rIL-4和rGM-CSF体外诱导骨髓前体细胞,培养第4天始加入红活麻乙酸乙酯有效部位20 mg/L处理,观察DC生长分化情况。用流式细胞仪检测DC表面抗原和共刺激因子,噻唑蓝(MTT)法检测经药物处理的DC体外刺激同种T细胞增殖活性,酶联免疫吸附法(ELISA)检测细胞培养上清液中IL-12p70分泌水平。结果红活麻乙酸乙酯有效部位抑制骨髓来源DC MHCⅡ和共刺激分子CD86的表达以及上清液中IL-12p70的分泌,与未经任何处理的正常DC比较差异有统计学意义。药物处理的DC与同种T细胞以不同比例混合培养时,刺激同种T细胞增殖能力显著下降。结论红活麻乙酸乙酯有效部位使小鼠骨髓来源树突状细胞处于未成熟状态,对其免疫学功能起负性调节作用,其免疫抑制活性的确定也为抗移植排斥反应药物的研发提供了方向。

Objective To investigate the effects of the acetoacetate active component of Hubei ethnodrug Honghuoma on differentiation and function of bone marrow-derived dendritic ceils （BM-DCs） in vitro. Methods BM-DCs were cultured in the medium containing rGM-CSF and rIL-4,with 20 mg/L acetoacetate extracts of Honghuoma. The immunophenotype of DCs in control and treated groups was analyzed by flow cytometry. The stimulatory activity of DCs on allogeneic T cells was tested by mixed lymphocyte reaction （MLR）. IL-12p70 production in culture supematant was examined by ELISA assay. Results Treatment with the acetoacetate active component of Honghuoma marked/y inhibited the expression of DCs surface antigen MHC I1 and costimulatory molecule CD86 ,and the secretion of IL-12p70 from DCs was decreased by the active component of Honghuoma as well. The treated DCs exhibited low stimulatory activity on allogeneic T cells when co-cultured at various rate of DC/T. Conclusion The acetoacetate active component of Honghuoma caused murine BM-DCs immaturity and exerted a negative effect on the immunologic functions of DCs in vitro.