不同浓度小檗碱对OX-LDL诱导单核细胞源性树突状细胞分化成熟的影响

Effects of different concentrations of berberine on differentiation and maturation of human monocyte-derived dendritic cells induced by oxidized low-density lipoprotein

目的探讨不同浓度小檗碱对氧化低密度脂蛋白(OX-LDL)诱导的人单核细胞源性树突状细胞(MoDCs)分化成熟的影响。方法采用免疫磁珠法分离正常人外周血CD14^+单核细胞,经重组人粒-单核细胞集落刺激因子和重组人白介素4诱导分化为Mo-DCs。将Mo-DCs随机分为实验1组、实验2组、实验3组、模型组和对照组。实验1、2、3组Mo-DCs加入终浓度为50 ng/mL的OX-LDL后分别加入终浓度为0.25、0.75、1.5μmol/L的小檗碱。模型组加入50 ng/mL的OX-LDL。对照组加入等量PBS。继续培养48 h。采用流式细胞术检测各组MoDCs表型(CD80、CD40、CD11c和HLA-DR),采用酶联免疫吸附法检测各组Mo-DCs细胞培养上清中白细胞介素12(IL-12)和白细胞介素10(IL-10)水平。结果与对照组相比,模型组Mo-DCs表面分子CD80、CD40和HLA-DR的表达均较高(P均<0.05),细胞培养上清液IL-12水平较高(P<0.05),IL-10水平较低(P<0.05)。与模型组比较,实验1、2、3组Mo-DCs表面分子CD80、CD40和HLA-DR的表达均较低(P均<0.05),细胞培养上清液IL-12水平较低(P均<0.05),IL-10水平较高(P均<0.05)。与实验1组比较,实验2、3组Mo-DCs表面分子CD80、CD40和HLA-DR的表达均较低(P均<0.05),细胞培养上清液IL-12水平较低(P均<0.05),IL-10水平较高(P均<0.05)。与实验2组比较,实验3组Mo-DCs表面分子CD80、CD40和HLA-DR的表达均较低(P均<0.05),细胞培养上清液IL-12水平较低(P<0.05),IL-10水平较高(P<0.05)。结论小檗碱可阻碍OX-LDL诱导的单核细胞源性树突状细胞免疫成熟,此效果有剂量依赖性。

Objective To investigate the influence of different concentrations of berberine on the differentiation and maturation of human monocyte-derived dendritic cells （Mo-DCs） induced by oxidized low-density lipoprotein （OX-LDL）.Methods Human peripheral blood CD14＋ monocytes were separated by magnetic activated cell sorting and were induced to differentiate into Mo-DCs by the recombinant human granulocyte-monocyte colony-stimulating factor and recombinant human interleukin-4.Mo-DCs were randomly divided into the experimental groups 1, 2, 3, the model group, and control group.After being added with OX-LDL at the final concentration of 50 ng/mL, Mo-DCs in the experimental groups 1, 2, and 3 were added with berberine at the final concentrations of 0.25, 0.75, and 1.5 μmol/L, respectively.The Mo-DCs in the model group were added with 50 ng/mL OX-LDL.Mo-DCs in the control group were given the same amount of PBS.They were all cultured for 48 h.The phenotypes of Mo-DCs （CD80, CD40, CD11c and HLA-DR） were detected by flow cytometry.The concentrations of interleukin-12 （IL-12） and interleukin-10 （IL-10） in the supernatant of Mo-DCs were detected by ELISA.Results Compared with the control group, the expression of CD80, CD40, and HLA-DR was significantly higher （all P〈0.05）;the concentration of IL-12 in Mo-DCs was higher （P〈0.05） whereas the concentration of IL-10 was lower in the model group （P〈0.05）.Compared with the model group, the expression of CD80, CD40, and HLA-DR was significantly lower （all P〈0.05）;the concentration of IL-10 in Mo-DCs was higher whereas the concentration of IL-12 was lower in the all experimental groups （P〈0.05）.Compared with the experimental group 1, the expression of CD80, CD40, and HLA-DR was significantly lower （all P〈0.05）;the secretion of IL-10 on Mo-DCs was higher whereas IL-12 was lower in the experimental groups 2 and 3 （P〈0.05）.Compared with experimental group 2, the expression of CD80, CD40, and HLA-DR was significantly lower （all P〈0.05）;the concentration of IL-10 in Mo-DCs was higher whereas the concentration of IL-12 was lower in the experimental group 3 （P〈0.05）.Conclusion Berberine inhibits the immune maturation of human Mo-DCs induced by OX-LDL and this effect is dose dependent.