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为了调查华蟾毒精(CBG)对树突状细胞分泌功能的影响,采用小鼠粒细胞巨噬细胞集落刺激因子(GM-NSF)和IL-4体外联合诱导培养小鼠骨髓细胞,获得树突状细胞(DCs)。经免疫磁珠分选法纯化获取CDllc+树突状细胞。试验组CDllc+树突状细胞中加入不同浓度CBG,空白对照组加等量RPMI-1640,阳性对照组加LPS。作用48 h后,采用酶联免疫吸附法(ELISA)检测培养上清液中细胞因子IL-12p70、IL-10及IL-1β含量。结果表明:在适量浓度10~50μg/mL范围内,CBG均以剂量依赖的方式上调DCs分泌IL-12p70和IL-1β水平,下调IL-10的分泌水平。说明CBG能够通过调控DCs分泌Th1、Th2型细胞因子的分泌,来诱导Th1型细胞免疫应答。
In order to investigate the effect of bufalin (CBG) on the secretory function of dendritic cells, the mouse bone marrow cells were induced by the combination of GM-CSF and IL-4 in vitro. Dendritic cells (DCs). CD11c + dendritic cells were obtained by immunomagnetic bead sorting. Experimental group CD11c + dendritic cells added with different concentrations of CBG, blank control group with equal amount of RPMI-1640, positive control group plus LPS. After 48 hours, the contents of cytokines IL-12p70, IL-10 and IL-1βwere detected by enzyme-linked immunosorbent assay (ELISA). The results showed that CBG up-regulated the levels of IL-12p70 and IL-1β secreted by DCs and down-regulated the secretion of IL-10 in a dose-dependent manner in the range of 10-50 μg / mL. CBG can regulate the secretion of Th1, Th2-type cytokines secreted by DCs to induce Th1-type cellular immune response.