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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
Objective: To investigate the effect of NFκB on the induction of iNOS gene expression in rat pulmonary microvascular endothelial cells and the effect of antioxidants on iNOS induced expression of endothelial cells. Methods: The level of NO-2 in cell culture supernatant was measured by Griess method to reflect the production of NO. The iNOS mRNA level was analyzed by Northern blotting. The binding activity of NFκB in nucleus was detected by EMSA. RESULTS: Antioxidants could block NO production and iNOS mRNA expression induced by LPS and TNFα in endothelial cell culture system. LPS and TNFα induced NFκB activation in rat pulmonary microvascular endothelial cells and could be blocked by antioxidants. CONCLUSION: The expression of iNOS gene in endothelial cells induced by LPS and TNFα depends on the activation of NFκB. Antioxidants can block the induction of iNOS gene expression by inhibiting the activation of NFκB