miR-155在3-oxo-C12-HSL阻碍人单核细胞源性树突状细胞成熟过程中的表达变化

Change in miR-155 expression during the 3-oxo-C12-HSL inhibition of maturation of human monocyte-derived dendritic cells

目的探讨miR-155在N-3-氧代十二烷酰-L-同型丝氨酸内酯(3-oxo-C12-HSL)阻碍人单核细胞源性树突状细胞(Mo-DCs)成熟过程中的表达变化。方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IL-4诱导分化为Mo-DCs。将不成熟Mo-DCs分为阴性对照组、模型组、实验组1、实验组2,阴性对照组加入同体积PBS,模型组加入终浓度为1μg/m L的LPS,实验组1和实验组2分别加入终浓度为1μg/m L的LPS后分别加入终浓度为50μmol/L和100μmol/L的3-oxo-C12-HSL,处理48 h。采用流式细胞术检测各组Mo-DCs表型(CD80、CD40、CD11c和HLA-DR),酶联免疫吸附法检测细胞培养上清细胞因子(IL-12、IL-10)水平,荧光定量PCR检测miR-155表达。结果与阴性对照组相比,模型组CD80、CD40、HLA-DR表达高,IL-10水平低,IL-12水平高,miR-155表达高(P均<0.05)。实验组1、实验组2与模型组相比,实验组2与实验组1相比,CD80、CD40、HLA-DR表达低,IL-10水平高,IL-12水平低,miR-155表达低(P均<0.05)。结论 miR-155在3-oxo-C12-HSL阻碍人Mo-DCs成熟过程中表达降低,3-oxo-C12-HSL可能通过降低miR-155表达阻碍Mo-DCs成熟。

Objective To investigate the expression changes of miR-155 during the 3-oxo-C12-HSL inhibition of maturation of human monocyte-derived dendritic cells(Mo-DCs).Methods Human peripheral blood CD14+monocytes were isolated and induced into Mo-DCs by recombinant human granulocyte-mononuclear colony-stimulating factor and recombinant human interleukin-4.Immature Mo-DCs were divided into the negative control group,model group,experimental group 1,and experimental group 2.The negative control group was added with the same volume of PBS,the model group was added with LPS(1μg/mL),the experimental group 1 was added with LPS(final concentration of 1μg/mL)and 3-oxo-C12-HSL with a final concentration of 50μmol/L,the experimental group 2 was added with LPS(1μg/mL)and 3-oxo-C12-HSL with a final concentration of 100μmol/L for 48 h.The phenotypes(CD80,CD40,CD11c,and HLA-DR)of Mo-DCs in each group were detected by flow cytometry.The levels of cytokines(IL-12 and IL-10)in the cell culture supermatants were detected by enzyme-linked immunosorbent assay.The expression of miR-155 was detected by real-time fluorescent quantitative PCR.Results Compared with the negative control group,the expression of CD80,CD40,and HLA-DR in the model group was higher,the IL-10 level was lower,the IL-12 level was higher,and the expression of miR-155 was higher(P<0.05).When we compared the experimental group 1 and experimental group 2 with the model group,as well as experimental group 2 with experimental group 1,the expression of CD80,CD40,and HLA-DR was lower,the IL-10 level was higher,the IL-12 level was lower,and the miR-155 expression was lower(all P<0.05).Conclusion The expression of miR-155 during the 3-oxo-C12-HSL inhibition of maturation of Mo-DCs is decreased,suggesting that 3-O-C12-HSL may inhibit the maturation of Mo-DCs by reducing the expression of miR-155.