黄芪多糖诱导脐血单个核细胞向树突状细胞分化的免疫机制研究

Study on Immunologic Mechanisms of Astragalus polysaccharides Inducing the Human Cord Blood Monocyte to Dendritic Cell

目的:探讨黄芪多糖诱导脐血来源的单个核细胞分化为树突状细胞的免疫学机制。方法:采集新鲜脐血,Ficoll淋巴细胞分离液梯度离心法获得单个核细胞,对照组以含10%胎牛血清的RPMI-1640完全培养基进行培养,实验组加APS(100mg.L-1)进行诱导;两组细胞隔天半量换液,实验组补充半量的APS;酶联免疫吸附法(ELISA)检测培养至第4、6、8、10天培养液上清中GM-CSF、TNF-α、IL-10的浓度。结果:实验组GM-CSF、TNF-α的浓度在第4、6、8、10天均明显高于对照组,差异具有统计学意义(P<0.05);负调控因子IL-10的浓度,实验组亦高于对照组(P<0.05);将TNF-α与IL-10相对应的每个标本的四个复孔值进行两两相比(TNF-α/IL-10),其比值以R表示,实验组与对照组之间差异性,且实验组R值均高于对照组,第6、8、10天差异具有显著性(P<0.01)。结论:黄芪多糖促使单个核细胞分泌GM-CSF、TNF-α诱导单个核细胞向树突状细胞分化、成熟;黄芪多糖诱导单个核细胞分泌IL-10,抑制其向树突状细胞分化成熟,但TNF-α与IL-10的比值R较高,且呈动态变化规律,抑制IL-10对树突状细胞的影响。

Objective：To explore the immunologic mechanisms how APS（APS） induced the cord blood monocyte in vitro to the dendritic cells（DCs）.Methods：Collected new cord blood,and isolated the mononuclear cells from cord blood with Ficoll-hypaque centrifugation method.The control group was cultured with RPMI-1640 com-culture medium including 10 per fetal calf serum.The APS group was cultured with APS（100mg L-1） another.Every two days changed half of the medium and the APS was added with the same density in the APS group.The density of GM-CSF、TNF-α and IL-10 in the cell culture fluid were detected with Enzyme-linked immunoadsordent assay（ELISA） in the day of 4th、6th、8th and 10th.Results：The density of GM-CSF、TNF-α were higher in the exp experiment group than that of in the control group.The differences were significance（P0.05）.The density of IL-10 was higher too in the experiment group（P0.05）.The result of TNF-α/IL-10 was named R,and the R value of the experiment group were higher than that in the control group in the four repeat-holes,the difference were significant in the 6th,8th and 10th days（P0.01）.Conclusion：APS can stimulate mononuclear cells to secrecte GM-CSF、TNF-α and induce the cord blood monocyte to committed differentiate into functional DCs.In this process,IL-10 was secrected too to prevent the cells changed into DCs,but this role was restrained by high level TNF-α.,and R value changed with time regularitively.