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〔目的〕用化学发光法检测培养细胞释放的 NO。〔方法〕NO2 -是生物样品中 NO代谢产物的标志物。在酸性条件下 ,用 KI将 NO2 - 还原成 NO,NO和 luminol- H2 O2 体系产生化学发光。按照去细胞、去蛋白、加络合剂 EDTA、将样品和 KI溶液充氮气、把 KI加到样品中、测化学发光值这几个步骤 ,我们检测了加脂多糖 (L PS)、加 N (G) -甲基 - L-精氨酸 (L- NMMA)、不加任何物质作对照三组培养细胞释放 NO的水平。〔结果〕NO水平由高到低依次为 :L PS组 >对照组 >L- NMMA组 ,在 1.0× 10 - 1 1 mol/L至 1.7× 10 - 9m ol/L 范围内线性良好 ,检出限为 1.0× 10 - 1 2mol/L。〔结论〕此方法可间接检测培养细胞释放的 NO,且灵敏度高。
[Purpose] The chemiluminescence method was used to detect NO released from cultured cells. [Method] NO2 - is a marker of NO metabolites in biological samples. Under acidic conditions, NO2 - is reduced to NO with KI, producing chemiluminescence with NO and luminol - H2 O2 systems. According to the steps of removing cells, removing protein, adding complexing agent EDTA, filling sample and KI solution with nitrogen gas, adding KI to the sample and measuring chemiluminescence value, we tested the effects of adding lipopolysaccharide (LPS), adding N (G) -methyl-L-arginine (L-NMMA) without any substance as control. [Results] The results showed that the levels of NO from high to low were as follows: L PS group> control group> L NMMA group, the linearity was good in the range of 1.0 × 10 - 1 1 mol / L to 1.7 × 10 - 9 mol / L, The limit is 1.0 × 10 - 1 2mol / L. [Conclusion] This method can indirectly detect the NO release from cultured cells with high sensitivity.