瑞香素对人PBMC的细胞因子和TLRs mRNA表达的影响
Effects of daphnetin on the mRNA expression of cytokines and TLR1- 10 in PBMC
曾小平 1姜青龙 2傅颖媛 1况南珍 1尧荣凤1
作者信息
- 1. 330006,南昌大学基础医学院免疫学教研室
- 2. 330006南昌,江西省临床检验中心
- 折叠
摘要
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、1FN-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.
Abstract
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 T 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.
关键词
瑞香素/淋巴细胞/Toll样受体/实时荧光定量PCRKey words
Daphetin/Lymphocyte/Toll-like receptor/Real time-PCR引用本文复制引用
基金项目
江西省自然科学基金(2008GZY0031)
出版年
2012
NETL
NSTL
维普
万方数据