DIRECT DETECTION OF TISSUE NITRIC OXIDE IN SEPTIC MICE

尽管一氧化氮(NO)是一种中枢调节因子 内毒素诱导的脓毒症，直接检测体内组织中的一氧化氮，有 直到最近还很困难，而且技术一直依赖于间接 血液或侵入性技术中副产物的测量。我们有 利用电子顺磁共振(EPR)与 自旋捕获技术，直接、非侵入性地检测NO， 从败血症小鼠的组织中提取。相对信号强度上升 从NO与铁和二乙基二硫代氨基甲酸酯(DETC)络合的测定 直接从肝脏和肾脏给予内毒素的小鼠最大 在内毒素后6小时。获得的EPR信号的质量 (高信噪比为15：1)使用我们的实验设置和 L乐队的EPR硬件是这样的，我们能够建立一个时间 内毒素在组织中产生一氧化氮的过程及其测定 来自其他器官(肾和脾)的NO。这项技术是 扩展到我们第一次植入氧气的实验 敏感物质(Gloxy)进入小鼠肝脏，然后进行监测 内毒素过后没有产生。由于EPR Gloxy的光谱和NO-Fe-(DETC)2的光谱不重叠，我们是 能够同时监测组织中NO的产生和PO2，在 实时的。

Although nitric oxide (NO) is a central mediator during endotoxin-induced sepsis, direct detection of tissue NO in vivo, has until recently been difficult, and techniques have relied on indirect measurement of byproducts in blood or invasive technology. We have utilized electron paramagnetic resonance (EPR) in conjunction with the spin-trapping technique to detect NO directly, and non-invasively, from the tissue of septic mice. Relative signal intensity arising from NO complexed with iron and diethyldithiocarbamate (DETC) measured directly from the liver and kidney of mice given endotoxin was maximal at 6 hours post endotoxin. The quality of the EPR signal obtained (high signal to noise ratio of 15:1) using our experimental set-up and L-band EPR hardware was such that we were able to establish a time course of NO production in tissue following endotoxin, and measurement of NO from other organs (kidney and spleen). This technique was extended to experiments in which we first implanted an oxygen sensitive material (gloxy) into the liver of mice, and then monitored NO production following endotoxin. Due to the fact that the EPR spectrum from gloxy and that of NO-Fe-(DETC)2 do not overlap, we were able to monitor NO production and pO2 simulatneously in tissue, in real time.