Fujii H, Berliner L J
Department of Inflammation Research, Tokyo Metropolitan Institute of Medical Science, Rinshoken, Japan.
Phys Med Biol. 1998 Jul;43(7):1949-56. doi: 10.1088/0031-9155/43/7/016.
This paper discusses, compares and evaluates various in vivo EPR methods of detection of nitric oxide (NO). In particular the various classes of agents are: Fe(II)-dithiocarbamate derivative complexes of MGD (N-methyl-D-glucamine dithiocarbamate) and DTCS [N-(dithiocarboxy)sarcosine], stable imidazolineoxy N-oxides and nitronyl N-oxides, and NO-sensitive chars. As a specific example direct, real-time, in vivo measurements of nitric oxide (NO) in mice are described with the water soluble metal chelator complex (MGD)2-Fe(II), as monitored at L-band EPR. The three-line EPR spectrum of [(MGD)2-Fe(II)-NO] was observed non-invasively in both control animals injected with the preformed product [(MGD)2-Fe(II)-NO] and from lipopolysaccharide (LPS) treated mice subsequently injected with (MGD)2-Fe(II) complex. The [(MGD)2-Fe(II)-NO] spectrum was markedly suppressed after administration of phenyl N-tert-butyl nitrone (PBN) prior to LPS injection as PBN inhibits the expression of inducible nitric oxide synthase (iNOS). When 15N-arginine was administered to LPS-treated mice, an EPR spectrum consisting of both three- and two-line EPR signals (due to (MGD)2-Fe(II)-14NO and (MGD)2-Fe(II)-15NO respectively) was observed, confirming that the trapped NO was generated through the NOS enzyme and not other chemical routes.
本文讨论、比较并评估了检测一氧化氮（NO）的各种体内电子顺磁共振（EPR）方法。特别是各类试剂包括：MGD（N-甲基-D-葡糖胺二硫代氨基甲酸盐）和DTCS [N-（二硫代羧基）肌氨酸]的Fe（II）-二硫代氨基甲酸盐衍生物配合物、稳定的咪唑啉氧基N-氧化物和硝酰基N-氧化物，以及对NO敏感的炭。作为一个具体例子，描述了使用水溶性金属螯合剂配合物（MGD）2-Fe（II）在L波段EPR监测下对小鼠体内一氧化氮（NO）进行直接、实时的测量。在注射预制产物[（MGD）2-Fe（II）-NO]的对照动物以及随后注射（MGD）2-Fe（II）配合物的脂多糖（LPS）处理的小鼠中，均非侵入性地观察到了[（MGD）2-Fe（II）-NO]的三线EPR谱。在LPS注射前给予苯基N-叔丁基硝酮（PBN）后，[（MGD）2-Fe（II）-NO]谱明显受到抑制，因为PBN抑制诱导型一氧化氮合酶（iNOS）的表达。当向LPS处理的小鼠施用15N-精氨酸时，观察到由三线和二线EPR信号组成的EPR谱（分别归因于（MGD）2-Fe（II）-14NO和（MGD）2-Fe（II）-15NO），证实捕获的NO是通过NOS酶产生的，而非其他化学途径。
