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目的 :探索一氧化氮供体硝普钠 (SNP)在任洛氏溶液 (RL S)中 ,一氧化氮 (NO)释放通路及其动力学特征 ,为体外 NO定量生物学研究提供科学依据。方法 :在 37℃恒温避光条件下 ,对 NO特异氧化产物 NO2 - Griess还原显色 ,采用分光光度法 ,比较两种活性巯基化合物半胱氨酸 (CYS)和谷胱甘肽 (GSH)存在的 RL S中 ,SNP释放 NO的情况 (n=3)。结果 :无活性巯基存在的 RLS中未测得 NO;而当有 2 5 mmol/ L CYS存在时 ,SNP释放 NO呈一级动力学 ,速率常数 k=0 .1 397min- 1(r=0 .932 1 ) ,稳态浓度为 0 .87mmol/ L ,CYS浓度的平方与 SNP释放 NO呈线性关系 ;有2 5 mmol/ L GSH存在时 ,NO释放通路反应呈多相性 ,约在 1 0 min时 ,出现 0 .5 2 mmol/ L的峰值 ,后迅速下降 ,在 30 min时渐成 0 .1 8mmol/ L 的稳态。结论 :恒温避光 RL S中 ,SNP释放 NO呈 -SH依赖性。在 CYS体系中 ,NO释放呈一级动力学 ;在一定范围内改变 CYS浓度 ,对 NO释放影响较小。在 GSH体系中 ,30 min内 NO释放呈多相反应。
OBJECTIVE: To explore the nitric oxide (NO) release pathway and its kinetic characteristics of nitric oxide donor sodium nitroprusside (SNP) in Ringer’s solution (RL S), and provide a scientific basis for the quantitative bioassay of NO in vitro. Methods: NO2 - Griess reduction of NO was carried out under the condition of constant temperature and light at 37 ℃. Spectrophotometry was used to compare the existence of two active sulfhydryl compounds cysteine (CYS) and glutathione (GSH) Of RL S, SNP release NO (n = 3). RESULTS: NO was not detected in the presence of inactive sulfhydryl in the presence of RLS. When 25 mmol / L CYS was present, NO released by SNP showed a first order kinetics with a rate constant of k = 0.1397 min-1 (r = 932 1). The steady-state concentration was 0.87 mmol / L, and the square of CYS concentration was linear with SNP release. In the presence of 25 mmol / L GSH, NO release pathway reaction was heterogeneous, , The peak appeared at 0.52 mmol / L, then decreased rapidly and gradually became 0.18 mmol / L at 30 min. CONCLUSION: SNP released by SNP showed -SH-dependent in constant temperature darkness. In the CYS system, NO release was in first order kinetics. Changing CYS concentration within a certain range had little effect on NO release. In the GSH system, NO release was heterogeneous within 30 min.