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目的观察瑞香素对人外周血单个核细胞的细胞因子和Toll样受体表达的影响,研究瑞香素增强淋巴细胞功能的作用机制。方法分离人外周血T、B细胞和单核细胞,分别加入瑞香素,37℃,5%CO2培养48 h后,采用实时荧光定量PCR检测瑞香素对T细胞IL-2、IFN-γ,B细胞IL-12,单核细胞IL-1 mRNA及T、B细胞TLR1～10的mRNA表达的影响;同时检测先用抗TLR4单抗封闭,再用瑞香素处理细胞后上述细胞因子和TLRs的mRNA表达变化。结果瑞香素能明显上调T细胞IL-2及IFN-γ、B细胞IL-12、单核细胞IL-1 mRNA和T细胞TLR1、TLR4,B细胞TLR4、TLR9 mRNA的表达(P﹤0.05),其中T细胞IL-2、B细胞IL-12、单核细胞IL-1 mRNA的表达和T细胞TLR4,B细胞TLR4、TLR9 mRNA表达可被抗TLR4单抗部分阻断。结论瑞香素增强淋巴细胞的免疫功能的可能机制是通过上调T细胞TLR1、TLR4,B细胞TLR4、TLR9的表达,分别参与其细胞内信号转导,从基因转录水平促进T、B细胞分泌细胞因子。
Objective To observe the effects of daphnetin on the expression of cytokines and Toll-like receptors in human peripheral blood mononuclear cells and to study the mechanism of daphnetin in enhancing lymphocyte function. Methods T cells, monocytes and peripheral blood mononuclear cells were isolated from human peripheral blood mononuclear cells (PBMCs). Daphnetin, 37 ℃ and 5% CO2 were added respectively for 48 hours. The changes of IL-2, IFN- IL-12, monocyte IL-1 mRNA and T, B cells TLR1 ~ 10 mRNA expression; simultaneous detection of anti-TLR4 monoclonal antibody blocking, and then daphnetin treatment of cells after the above cytokines and TLRs mRNA Change of expression Results Daphnetin significantly up-regulated the expressions of IL-2 and IFN-γ, IL-12, IL-1 and TLR4, TLR9 mRNA in T cells (P <0.05) The expression of IL-2, IL-12 and monocyte IL-1 mRNA in T cells and TLR4 in T cells and TLR4 mRNA in B cells were partially blocked by anti-TLR4 mAb. Conclusion Daphnetin may enhance the immune function of lymphocytes by up-regulating the expression of TLR4, TLR9 in TLR1, TLR4 and B cells, and participate in their intracellular signal transduction respectively, and promote the secretion of cytokines from T and B cells .