巨噬细胞中诱导型一氧化氮合酶来源的NO对共培养HL60 细胞凋亡的影响

 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。

Effects of Inducible Nitric Oxide Synthase Derived Nitric Oxide on Apoptosis of HL60 Cells Co-cultured with RAW264.7 Macrophages

Objective 　To study effects of inducible nitric oxide synthase (iNOS)-derived nitric oxide (NO) on apoptosis of HL60 cells co2cultured with RAW 264. 7 macrophages. Methods 　Upon stimulation with lipopolysaccharide (LPS) and interferon-γ( IFN-γ) , inducible nitric oxide synthase gene was expressed in RAW 264. 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 cytomet ry (FCM) , transmission elect ron microscopy ( TEM) and DNA agarose gelelect rophoresis. Results 　The results showed that iNOS-derived nitric oxide caused oxidative damage of HL60 cells co-cultured with RAW 264. 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 f ragmentation. Conclusion 　The result s suggested important effect of iNOS-derived nitric oxide on apoptosis of cells in RAW 264. 7 macrophages.