黄芪多糖通过NF-κB诱导巨噬细胞产生NO和TNF-α  

The role of nuclear factor-kappaB in the induction of nitric oxide and tumor necrosis factor-alpha by Astragalus mongholicus polysaccharides in macrophages

作　　者：朱杰[1] 肖震[1] 沈月爽[1] 吴国友[1] 

机构地区：[1]浙江医院检验科,杭州310013

出　　处：《中华微生物学和免疫学杂志》2010年第6期511-515,共5页Chinese Journal of Microbiology and Immunology

摘　　要：目的 通过体外试验研究黄芪多糖（Astragalus mongholicus polysaccharides,ASP）激活巨噬细胞产生NO和TNF-α的分子机制和细胞内信号转导机制.方法 黄芪多糖刺激RAW264.7细胞,用Western blot方法 检测细胞核内核转录因子-κB（NF-κB）的变化.用Griess还原法观察黄芪多糖对巨噬细胞释放NO的作用的影响以及NF-κB抑制剂对黄芪多糖诱导巨噬细胞释放NO作用和分泌TNF-α的影响.ELISA法检测黄芪多糖对巨噬细胞分泌肿瘤坏死因子-α（TNF-α）的变化.结果 100μg/ml黄芪多糖刺激RAW264.7细胞,4 h后可引起细胞核内NF-κB含量显著增加,6 h达到顶峰.16 h后可显著诱导NO[（18.9±1.5）μmol/L;P〈0.01]释放和TNF-α分泌[（81.2±16.7）pg/ml,P〈0.01]的增加,以及诱导型一氧化氮合酶（iNOS）[（23.54±2.41）U/mg蛋白质,P〈0.01]活性的增加.NF-κB抑制剂吡咯烷二硫代氨基甲酸盐（PDTC）可明显抑制黄芪多糖诱导RAW264.7生成NO和分泌TNF-α.结论 NF-κB在黄芪多糖诱导巨噬细胞生成NO和TNF-α过程中发挥重要作用.Objective To explore the molecular and cell signal transduction mechanism of Astragalus mongholicus polysaccharides （ASP） on macrophage. Methods After stimulating RAW264.7, the change in value of NF-κB was determined by Western blot. The induction of NO and secretion of TNF-α by ASP in macrophage was observed with or without inhibitor of NF-κB using Griess method. Moreover, protein levels of TNF-α secreted by macrophage were investigated with ELISA in respond to ASP. Results 4 h after stimulation by 100 μg/ml ASP, the concentration of NF-κB in nucleus increased significantly, peaked at 6 h. 16 h after stimulation by 100 μg/ml ASP, the activity of iNOS[（23.54±2.41） U/mg protein; P〈0.01], producton of NO [（18.9±1.5）μmol/L, P〈0.01] and level of TNF-α[（81.2±16.7）pg/ml, P〈0.0l] in macrophage were improved markedly. Blocking NF-κB with inhibitor results in decreased levels of NO and TNF-α. Conclusion The results suggest that NF-κB play an important role in induction of NO and TNF-α by ASP in macrophage.

关 键 词：黄芪多糖 巨噬细胞 核转录因子-ΚB 

分 类 号：R285.5[医药卫生—中药学]