摘要
〔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。
睴bjective〕TO detect the NO released from Cultured cells by chemiluminescent method.〔Methods〕Nitrite ion(NO 2 -) can be taken as a marker of nitric oxide(NO) production in biological sample NO 2 - can be reduced to NO by potassium iodide(KI) in acidic solution NO react with luminol-H 2O 2 system to form chemiluminescence The NO levels of three groups of cultured cells which were cultured respectively in medium 1640 with lipopolysaccharide (LPS),medium 1640 with L-NMMA,and medium 1640 as control were detected The CL emission from cultured cells can be detected as follows:remove cells→remove protein→add EDTA to sample→bubbled N 2 in sample and fresh KI solution→add KI solution to sample→detection of light emission 〔Results〕the logarithm value of relative light intensity is proportional to that of concentration of NO in the range from 1 0×10 -11 to 1 7×10 -9 mol/L,with a detection limit of 1 7×10 -12 mol/L,the order of NO concentrations is LPS group>controlled group>L-NMMA group 〔Conclusions〕This method can assay the NO released from cultured cells indirectly,and the sensitivity is high.
出处
《中国卫生检验杂志》
CAS
2000年第3期277-279,共3页
Chinese Journal of Health Laboratory Technology
基金
国家自然科学基金!资助课题 (编号 :39670 2 19)
关键词
一氧化氮
培养细胞
化学发光
nitric oxide
cultured cells
chemiluminescence
