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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 investigate the immunological mechanism of astragalus polysaccharides in inducing dendritic cells derived from cord blood mononuclear cells. Methods: Mononuclear cells were obtained by fresh cord blood and Ficoll lymphocyte separation liquid gradient centrifugation. The control group was cultured in RPMI-1640 complete medium containing 10% fetal bovine serum. The experimental group received APS (100 mg.L-1) The amount of APS in the experimental group was supplemented with half the amount of APS. The levels of GM-CSF and TNF-α in the culture supernatant of the 4th, 6th, 8th and 10th days after culture were detected by enzyme-linked immunosorbent assay (ELISA) α, IL-10 concentrations. Results: The concentrations of GM-CSF and TNF-α in the experimental group were significantly higher than those in the control group on the 4th, 6th, 8th and 10th days (P <0.05). The concentrations of negative regulatory factor IL- The experimental group was also higher than that of the control group (P <0.05). The four replicate values of each specimen corresponding to TNF-α and IL-10 were compared with each other (TNF-α / IL-10) R, the difference between the experimental group and the control group, and the experimental group R values were higher than the control group, 6,8,10 days difference was significant (P <0.01). Conclusion: Astragalus polysaccharide can induce the monocyte to secrete GM-CSF, TNF-αinduces the differentiation and maturation of mononuclear cells to dendritic cells. Astragalus polysaccharide can induce mononuclear cells to secrete IL-10, inhibit the dendritic cells to differentiate and mature, However, the ratio R of TNF-α and IL-10 was high and showed a dynamic change rule, which inhibited the effect of IL-10 on dendritic cells.