中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2010年
6期
511-515
,共5页
朱杰%肖震%沈月爽%吴国友
硃傑%肖震%瀋月爽%吳國友
주걸%초진%침월상%오국우
黄芪多糖%巨噬细胞%核转录因子-κB
黃芪多糖%巨噬細胞%覈轉錄因子-κB
황기다당%거서세포%핵전록인자-κB
Astragalus mongholicus polysaccharides%Macrophage%NF-κB
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.
目的 通過體外試驗研究黃芪多糖(Astragalus mongholicus polysaccharides,ASP)激活巨噬細胞產生NO和TNF-α的分子機製和細胞內信號轉導機製.方法 黃芪多糖刺激RAW264.7細胞,用Western blot方法 檢測細胞覈內覈轉錄因子-κB(NF-κB)的變化.用Griess還原法觀察黃芪多糖對巨噬細胞釋放NO的作用的影響以及NF-κB抑製劑對黃芪多糖誘導巨噬細胞釋放NO作用和分泌TNF-α的影響.ELISA法檢測黃芪多糖對巨噬細胞分泌腫瘤壞死因子-α(TNF-α)的變化.結果 100μg/ml黃芪多糖刺激RAW264.7細胞,4 h後可引起細胞覈內NF-κB含量顯著增加,6 h達到頂峰.16 h後可顯著誘導NO[(18.9±1.5)μmol/L;P<0.01]釋放和TNF-α分泌[(81.2±16.7)pg/ml,P<0.01]的增加,以及誘導型一氧化氮閤酶(iNOS)[(23.54±2.41)U/mg蛋白質,P<0.01]活性的增加.NF-κB抑製劑吡咯烷二硫代氨基甲痠鹽(PDTC)可明顯抑製黃芪多糖誘導RAW264.7生成NO和分泌TNF-α.結論 NF-κB在黃芪多糖誘導巨噬細胞生成NO和TNF-α過程中髮揮重要作用.
목적 통과체외시험연구황기다당(Astragalus mongholicus polysaccharides,ASP)격활거서세포산생NO화TNF-α적분자궤제화세포내신호전도궤제.방법 황기다당자격RAW264.7세포,용Western blot방법 검측세포핵내핵전록인자-κB(NF-κB)적변화.용Griess환원법관찰황기다당대거서세포석방NO적작용적영향이급NF-κB억제제대황기다당유도거서세포석방NO작용화분비TNF-α적영향.ELISA법검측황기다당대거서세포분비종류배사인자-α(TNF-α)적변화.결과 100μg/ml황기다당자격RAW264.7세포,4 h후가인기세포핵내NF-κB함량현저증가,6 h체도정봉.16 h후가현저유도NO[(18.9±1.5)μmol/L;P<0.01]석방화TNF-α분비[(81.2±16.7)pg/ml,P<0.01]적증가,이급유도형일양화담합매(iNOS)[(23.54±2.41)U/mg단백질,P<0.01]활성적증가.NF-κB억제제필각완이류대안기갑산염(PDTC)가명현억제황기다당유도RAW264.7생성NO화분비TNF-α.결론 NF-κB재황기다당유도거서세포생성NO화TNF-α과정중발휘중요작용.
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.
