瑞香素对人PBMC的细胞因子和TLRs mRNA表达的影响

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。

Effects of daphnetin on the mRNA expression of cytokines and TLR1-10 in PBMC

In this study,we will study the mechanism of daphnetin regulating lymphocytes by observing the effect of daphnetin on the expression of cytokine and Toll-like receptors(TLRs) in peripheral blood mononuclear cells(PBMC).T,B lymphocytes and monocytes separated from peripheral blood of human were respectively cultured with daphnetin at 37 ℃ for 48 hours.Then the effects of daphnetin on the mRNA expression of IL-2/IFN-γ in T cell,IL-12 in B cell,IL-1 in monocyte,as well as TLR1-10 in T and B cell were determined by the real-time fluorescence quantitative polymerase chain reaction(RT-PCR).Furthermore the mRNA of the cytokines and TLRs were tested again by RT-PCR after blocking the peripheral blood mononuclear cells(PBMC) with anti-TLR4.The results showed that daphnetin could increase the mRNA expression of IL-2/IFN-γ in T cell,IL-12 in B cell,IL-1 in monocyte,TLR1/TLR4 in T cell,and TLR4/TLR9 in B cell(P﹤0.05).Furthermore anti-TLR4 could partly block the unregulated mRNA expression of IL-2 in T cell,IL-12 in B cell,IL-1 in monocyte,TLR4 in T cell,TLR4/TLR9 in B cell(P﹤0.05).All this result indicate that daphnetin maybe upregulates the function of lymphocytes by increasing the expression of TLR1/TLR4 in T lymphocytes and TLR4/TLR9 in B lymphocytes which are concerned respectively with the intracellular signal transduction and the promotion of cytokines secretion of T and B lymphocytes.