Komarov A, Mattson D, Jones M M, Singh P K, Lai C S
Biophysics Research Institute, Medical College of Wisconsin, Milwaukee 53226.
Biochem Biophys Res Commun. 1993 Sep 30;195(3):1191-8. doi: 10.1006/bbrc.1993.2170.
We report here an in vivo spin-trapping technique combined with electron paramagnetic resonance (EPR) spectroscopy to measure nitric oxide (.NO) production in the blood circulation of conscious mice. The method is based on the trapping of nitric oxide (.NO) by a metal-chelator complex consisting of N-methyl-D-glucamine dithiocarbamate (MGD) and reduced iron (Fe2+) to form a stable and water-soluble [(MGD)2-Fe(2+)-NO] complex, which gives rise to a characteristic three-line EPR spectrum of a mononitrosyl-Fe2+ complex (aN = 12.5 G and g(iso) = 2.04) at ambient temperatures. After simultaneous intravenous injection of sodium nitroprusside (an .NO donor) and the [(MGD)2-Fe2+] complex into the lateral vein of the mouse tail, the appearance of the three-line spectrum in the blood circulation of the mouse tail was monitored continuously by using an S-band EPR spectrometer, operating at 3.5 GHz. This represents the first spin trapping of .NO in living animals.
我们在此报告一种体内自旋捕获技术,该技术结合电子顺磁共振(EPR)光谱来测量清醒小鼠血液循环中的一氧化氮(·NO)生成。该方法基于由N - 甲基 - D - 葡糖胺二硫代氨基甲酸盐(MGD)和还原铁(Fe2 +)组成的金属螯合物复合物捕获一氧化氮(·NO),形成稳定且水溶性的[(MGD)2 - Fe(2 +) - NO]复合物,在环境温度下会产生单亚硝基 - Fe2 +复合物的特征三线EPR光谱(aN = 12.5 G,g(iso) = 2.04)。在将硝普钠(一种·NO供体)和[(MGD)2 - Fe2 +]复合物同时静脉注射到小鼠尾巴的侧静脉后,使用工作在3.5 GHz的S波段EPR光谱仪连续监测小鼠尾巴血液循环中三线光谱的出现。这代表了在活体动物中首次对·NO进行自旋捕获。