参附注射液联合细胞因子对急性髓性白血病HL-60细胞来源的树突状细胞诱导生成及功能的影响

Effect of Shenfu injection with cytokines on the maturation and function of dendritic cells derived from AML HL-60 cells

目的研究参附注射液(SFI)联合细胞因子对HL-60细胞体外诱导树突状细胞(DCs)生成及功能的影响。方法用不同浓度的SFI、GM-CSF+IL-4及GM-CSF+IL-4联合不同浓度SFI三种组合在体外诱导培养HL-60细胞株获得DCs。观察DCs的形态变化、表型的表达率并鉴定DCs刺激T淋巴细胞的增殖能力,ELISA法测定上清液中INF-γ的含量。结果单用SFI不能诱导HL-60细胞分化为DCs。单用细胞因子GM-CSF/IL-4可诱导其分化为DCs,但不成熟。而不同浓度的SFI联合GM-CSF/IL-4对DCs有促进成熟和抑制增殖的双向调节作用。中浓度诱生的DCs细胞表型的表达率、刺激T淋巴细胞的增殖能力、培养上清液中INF-γ的含量均明显高于其余试验组(P<0.05)。结论适当浓度的SFI联合GM-CSF/IL-4在体外能更有效地诱导HL-60细胞分化为DCs,并促进DCs成熟,增强DCs功能。

Objective To investigate the effect of shenfu injection （SFI） with cytokines on the maturation and function of dendritic cells （DCs） derived from the HL-60 cells. Methods Different concentrations of SFI plus different concentrations of SFI or GM-CSF ＋ IL-4, or GM-CSF ＋ IL-4 were used to induce HL-60 cells getting DCs. The morphologic changes and the phenotypic expression rate were observed, at the same time, the capability of DCs stimulating T lymphocyte proliferation was tested. The level of INF-γ in supernatant was measured by ELISA method. Results Only SFI could not induce the HL-60 to derive DCs. While DCs could derive from HL-60 cells co-cultured with GM-CSF/IL-4 but it was immature. However, different concentration of SFI plus GM-CSF/IL-4 not only induced maturation of HL-60 DCs but also restrained propagation of cells. Compared with other control groups, ceils cultured with middle concentration expressed higher levels of phenotypes, elicited significantly stronger abilities of stimulating T lymphocyte proliferation, and secreted larger amounts of INF-γ in supernatant. Conclusion Appropriate concentrations of SFI plus GM-CSF/IL-4 could effectively induce differentiation and maturation of HL-60 DCs,and enhance their immune stimulatory function.