黄芪多糖促进人树突状细胞体外分化成熟的实验研究

Experimental study on astragalus polysaccharides promoting the human dendritic cells maturation in vitro

目的:研究黄芪多糖(APS)对人树突状细胞(DCs)体外分化成熟及存活时间的影响。方法:健康志愿者外周血单个核细胞培养2 h获得贴壁的单核细胞,加入含rhGM-CSF(1000 U/mL)r、hIL-4(500 U/mL)的无血清培养基,实验组再加入3种浓度的APS(1μg/mL,10μg/mL,100μg/mL),对照组加入等体积生理盐水,一组细胞体外培养7 d,收集悬浮细胞获得普通DCs(Mo-DCs)及APS诱导的DCs(APS-DCs),流式细胞仪测定细胞免疫表型,同种异体混和淋巴细胞反应检测DCs的免疫激活功能,另外一组细胞体外连续培养14 d,光镜下观察细胞形态及存活时间。结果:APS能够显著促进DCs表面免疫分子HLA-DR、CD83、CD80、CD86、CD40、CD54的表达,APS-DCs较Mo-DCs具有更强的T细胞体外激活能力,并显著延长DCs的体外存活时间达2～3 d。结论:APS可以促进DCs的成熟和延长DCs的存活时间,从而发挥增强机体免疫力的功能。

AIM：To study the effects of astragalus polysaccharides（APS） on maturation and survival time of human dendritic cells（DCs） in vitro. METHODS：Peripheral blood mononuclear cells from healthy volunteers were cultured for 2 hours to acquire adherent monocytes.The monocytes were cultured with rhGM-CSF（1000 U/mL） and rhIL-4（500 U/mL） in serum-free medium.The experimental groups were added three concentrations of APS（1 μg/mL,10 μg/mL,100 μg/mL）,the control group were added equal volume of saline.One group of cells were cultured for 7 days and the cell suspension were collected to get normal DCs（Mo-DCs） or APS-induced DCs（APS-DCs）.The immunophenotyping was detected by flow cytometry,T cells stimulating activity of DCs were detected by allogeneic mixed lymphocyte reaction.Another group of cells were continuously cultured for 14 days and the cell morphology and survival time were observed under light microscope.RESULTS：APS could significantly promote the DCs surface HLA-DR,CD83,CD80,CD86,CD40,CD54 expression.APS-DCs compared with Mo-DCs had a stronger T-cell activation activity.APS significantly extended the DCs survival time up to 2-3 days in vitro.CONCLUSION：APS could promote DCs maturation and extend the survival time of DCs,which may enhance the immunity.