【摘要】 目的 :探讨NFκB在大鼠肺微血管内皮细胞iNOS基因诱导表达中的作用及抗氧化剂对内皮细胞iNOS诱导表达的影响及其机制。方法 :Griess法测定细胞培养上清液NO-2 水平以反映NO的生成 ,Northern印迹分析iNOSmRNA水平 ,EMSA法测定细胞核内NFκB的结合活性。结果 :抗氧化剂可阻断内皮细胞培养体系LPS和TNFα诱导的NO生成及iNOSmRNA的表达 ;LPS和TNFα可诱导大鼠肺微血管内皮细胞NFκB的激活并可被抗氧化剂阻断。结论 :LPS和TNFα诱导的内皮细胞iNOS基因表达依赖于NFκB的激活 ;抗氧化剂可通过抑制NFκB的活化而阻断iNOS基因的诱导表达更多还原

【Abstract】 AIM:To investigate the role of nuclear factor κB (NF κB) in the induction of iNOS gene by TNFα and LPS in endothelial cells and the effect of antioxidant on the induction of iNOS. METHODS:Nitrite was determined based on Griess reaction. iNOS mRNA was analyzed using Northern blot. NF κB in the cell nucleole was detected with electrophoretic mobility shift assay.RESULTS:(1)NO production and iNOS mRNA expression induced by LPS and TNFα was blocked by pyrrolidine dithiocarbamate(PDTC) or N-acetylcysteine(NAC). (2)LPS and TNFα triggered the activation and translocation of NF κB, and PDTC or NAC inhibited the activation of NF κB induced by LPS and TNFα.CONCLUSIONS:(1)The induction of iNOS gene by TNFα and LPS is dependent on the activation of NF κB.(2)Antioxidants may inhibit the induction of iNOS gene through the inhibition of NF κB activation.更多还原