【摘要】 目的研究巨噬细胞中诱导型一氧化氮合酶(iN-OS)来源的一氧化氮(NO)对共培养HL60细胞凋亡的影响。方法以脂多糖(LPS)和γ-干扰素(INF-γ)诱导RAW264.7巨噬细胞iNOS基因的表达产生过量NO为实验模型,通过噻唑蓝(MTT)试验、蛋白质印迹分析、荧光分析、流式细胞术(FCM)、透射电镜和DNA琼脂糖凝胶电泳等分析技术,观察NO对共培养的HL60细胞存活率、bcl-2和bax蛋白表达、Caspase-3活性和细胞凋亡的影响。结果RAW264.7巨噬细胞中iNOS来源的NO对共培养HL60细胞能造成氧化损伤,降低细胞的存活率;bcl-2表达明显下降,而bax表达增加;激活Caspase-3和促进DNA的降解。结论巨噬细胞中iNOS来源的NO在诱导细胞凋亡中发挥重要的作用。更多还原

【Abstract】 Objective To study effects of inducible nitric oxide synthase(iNOS)-derived nitric oxide(NO) on apoptosis of HL60 cells co-cultured with RAW264.7 macrophages.Methods Upon stimulation with lipopolysaccharide (LPS) and interferon-γ(IFN-γ), inducible nitric oxide synthase gene was expressed in RAW264.7 macrophages, which caused the consequent generation of nitric oxide. Effects of nitric oxide on HL60 cells viability, expression of bcl-2 and bax protein, activity of Caspase-3 and cell apoptosis were evaluated with MTT assay, Western blot analysis, fluorescence analysis, flow cytometry(FCM), transmission electron microscopy (TEM)and DNA agarose gel electrophoresis.Results The results showed that iNOS-derived nitric oxide caused oxidative damage of HL60 cells co-cultured with RAW264.7 macrophages, and decreased cell viability, and evidently reduced expression of bcl-2 and increased expression of bax, and induced activity of caspase-3 and DNA fragmentation.Conclusion The results suggested important effect of iNOS-derived nitric oxide on apoptosis of cells in RAW264.7 macrophages.更多还原