银杏叶提取物对内毒素诱导巨噬细胞核因子-κB活化及炎性细胞因子表达的调节  

Effect of EGb761 on Activation of Nuclear Factor-κB and Expression of Infla mmatory Cytokines in Lipopolysaccharide-Induced in Macrophages

作　　者：张申[1] 黄雪梅[1] 卫涛涛[2] 

机构地区：[1]怀化医学高等专科学校医学检验系,中国湖南怀化418000 [2]中国科学院生物物理研究所结构与分子生物学研究中心,中国北京100101

出　　处：《生命科学研究》2005年第4期336-340,共5页Life Science Research

基　　金：湖南省教育厅高校科研资助项目(04C050)

摘　　要：探讨银杏叶提取物(EGb761)对内毒素(LPS)诱导RAW264.7细胞核因子-κB(NF-κB)活化及炎性细胞因子基因表达的调节,为银杏叶提取物的临床运用提供理论依据.分别用LPS或EGb761+LPS处理体外培养的小鼠巨噬细胞系RAW264.7细胞,采用蛋白质印迹分析检测细胞中NF-κB活性,用逆转录-聚合酶链反应(RT-PCR)和酶联免疫吸附法(ELISA)检测细胞中TNF-α、IL-1β、IL-6mRNA和蛋白的表达.研究结果表明LPS组NF-κB活性和TNF-α、IL-1β、IL-6含量在刺激后2～12h明显高于正常对照组,而EGb761+LPS组NF-κB活性和TNF-α、IL-1β、IL-6含量均显著低于LPS组.结果提示LPS可诱导RAW264.7细胞NF-κB活化,导致TNF-α、IL-1β、IL-6基因表达增强,而EGb761能抑制NF-κB活化而调节TNF-α、IL-1β、IL-6基因的表达.To investigate activation of nuclear faetor-κB （NF-κB）and the expression of inflammatory eytokines in lipopolysaccharide-induced RAW264.7 cells stimulated with extract of Ginkgo biloba（EGb761）. RAW264. 7 cells, a murine macrophage cell line, cultured in vitro were given with LPS or EGb761 ＋ LPS. The NF-κB activity of RAW264.7 cells was tested by Western blotting analysis. The expression of TNF-α, IL-1β, IL-6 in RAW264.7 cells were measured by reverse transcription polymerase chain reaction techniques （RT-PCR） and enzyme-linked immuno sorbent assay （ELISA）. The results showed that the activity of NF-κB and the level of TNF-α, IL-1β, IL-6 significantly increased from 2 to 12 hour after LPS stimulation. Compared with LPS-stimulated group, both NF-κB activity and concentration of TNF-α, IL-1β, IL-6 were significantly lowered in EGb761 ＋ LPS group. The results suggested LPS might activate NF-κB in RAW264.7 cells and induce the increase of transcription and expression of TNF-α, IL-1β, IL-6 genes; EGb761 could inhibit the activation of NF-κB and regulate the expression of TNF-α, IL-1β and IL-6 genes in RAW264. 7 cells.

关 键 词：银杏叶提取物 巨噬细胞 脂多糖 核因子-ΚB 肿瘤坏死因子α 

分 类 号：Q786[生物学—分子生物学]