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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 observe the effect of ethyl acetate active fraction of Rhododendron L. on the function and expression of dendritic cells (DCs) cultured in vitro. Methods The bone marrow precursor cells were induced by rIL-4 and rGM-CSF in vitro. On the fourth day of culture, the effective fraction of ethyl acetate was added at 20 mg/L to observe the growth and differentiation of DCs. DC surface antigens and co-stimulatory factors were measured by flow cytometry, and thymidine blue (MTT) assay was used to detect the proliferation activity of allogeneic T cells stimulated by drug-treated DCs in vitro. The cell culture supernatants were detected by enzyme-linked immunosorbent assay (ELISA). IL-12p70 secretion levels. Results The ethyl acetate active fraction of red live anesthesia inhibited the expression of bone marrow-derived DC MHCII and co-stimulatory molecule CD86 as well as the secretion of IL-12p70 in the supernatant, which was significantly different from that of normal DC without any treatment. When the drug-treated DC was mixed with the same kind of T cells in different proportions, the ability to stimulate the proliferation of the same type of T cells was significantly decreased. Conclusion The ethyl acetate active fraction of red live anesthesia immortalizes mouse bone marrow-derived dendritic cells, negatively regulates its immunologic function, and its immunosuppressive activity is also determined for the development of anti-graft rejection drug research and development. Direction.