Egashira Toru, Takayama Fusako
Department of Pharmacology and Therapeutics, Oita Medical University, 1-1, Idaigaoka, Hasama-machi, Oita 879-5593, Japan.
Nihon Yakurigaku Zasshi. 2002 Oct;120(4):229-36. doi: 10.1254/fpj.120.229.
The detection of free radicals generated within the body may contribute to clarifying the pathophysiological role of free radicals in disease processes. As an appropriate procedure to examine the generation of free radicals in a biological system, electron spin resonance (ESR) has emerged as a powerful tool for detection and identification. A method for determination of oxygen radical scavenging activity using ESR and the spin trapping technique was developed. Oxygen radicals were trapped by 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) or alpha-phenyl-N-t-butylnitrone (PBN), and the DMPO or PBN spin adduct signal was measured quantitatively by an ESR spectrometer. The spin trapping method using ESR has also been reported for not only in vitro and ex vivo measurements but also in vivo measurements. In in vivo ESR, nitroxyl radical is being used as a spin trap well. ESR signal intensities of nitroxyl radical are measured after administration to animals and the signal decay rates of nitroxyl radical have reported to be influenced by various types of oxidative stress. With this method, it is possible to specify the type of radical or the location at which the free radicals are produced. The spin trapping method by in vivo ESR is an effective procedure for giving non-invasive measurements in animals. ESR imaging in the organs of live animals can also be obtained after injection of nitroxyl radicals as an imaging agent using ESR-computed tomography. In vivo ESR imaging has been established as a powerful technique for determining the spatial distribution of free radicals in living organs and tissues.
检测体内产生的自由基可能有助于阐明自由基在疾病过程中的病理生理作用。作为检测生物系统中自由基产生的一种合适方法,电子自旋共振(ESR)已成为一种强大的检测和识别工具。开发了一种利用ESR和自旋捕获技术测定氧自由基清除活性的方法。氧自由基被5,5-二甲基-1-吡咯啉-N-氧化物(DMPO)或α-苯基-N-叔丁基硝酮(PBN)捕获,然后用ESR光谱仪对DMPO或PBN自旋加合物信号进行定量测量。利用ESR的自旋捕获方法不仅已报道可用于体外和离体测量,还可用于体内测量。在体内ESR中,硝酰自由基也被用作一种良好的自旋捕获剂。将硝酰自由基给予动物后测量其ESR信号强度,并且据报道硝酰自由基的信号衰减率受各种类型氧化应激的影响。通过这种方法,可以确定自由基的类型或自由基产生的位置。体内ESR的自旋捕获方法是在动物中进行非侵入性测量的有效方法。在注射硝酰自由基作为成像剂后,还可以使用ESR计算机断层扫描获得活体动物器官的ESR成像。体内ESR成像已成为确定活器官和组织中自由基空间分布的强大技术。