Murrant C L, Reid M B
Department of Pharmacology and Physiology, University of Rochester, Rochester, New York 14642, USA.
Microsc Res Tech. 2001 Nov 15;55(4):236-48. doi: 10.1002/jemt.1173.
Reactive oxygen species (ROS) and reactive nitrogen species (RNS) are usually identified with pathological states and mediators of cellular injury. However, over the last decade ROS and RNS have been identified in skeletal muscle under physiological conditions. Detection of ROS and RNS production by skeletal muscle cells is fundamental to the problem of differentiating between physiological and pathological levels. The goal of this paper is to review the techniques that have been used to detect ROS and RNS in skeletal muscle. Electron spin resonance, fluorescent assays, cyotchrome c reduction, chemiluminescence, hydroxylation of salicylate, and nitration of phenylalanine are some of the assay systems that have been used thus far. A large body of evidence now indicates that ROS and RNS are continually produced by many different skeletal muscle types studied in vivo, in situ, and in vitro. Under resting conditions, ROS and RNS are detectable in both intracellular and extracellular compartments. Production increases during both non-fatiguing and fatiguing muscle contractions. In the absence of disease, the individual molecular species detected in skeletal muscle include parent radicals for the ROS and RNS cascades: superoxide anions and nitric oxide. Both are generated at rates estimated to range from pmol-to-nmol/mg muscle/minute. Evidence indicates that hydrogen peroxide, hydroxyl radicals, and peroxynitrite are also present under physiological conditions. However, the molecular species that mediate specific biological effects remains largely undetermined, as do the sources of ROS and RNS within muscle fibers. Eventual delineation of the mechanisms whereby ROS and RNS regulate cellular function will hinge on our understanding of the production and distribution of ROS and RNS within skeletal muscle.
活性氧（ROS）和活性氮（RNS）通常与病理状态以及细胞损伤介质相关联。然而，在过去十年中，人们已在生理条件下的骨骼肌中发现了ROS和RNS。检测骨骼肌细胞中ROS和RNS的产生对于区分生理水平和病理水平的问题至关重要。本文的目的是综述用于检测骨骼肌中ROS和RNS的技术。电子自旋共振、荧光测定、细胞色素c还原、化学发光、水杨酸羟化以及苯丙氨酸硝化是目前已使用的一些检测系统。现在大量证据表明，在体内、原位和体外研究的许多不同类型骨骼肌中持续产生ROS和RNS。在静息条件下，细胞内和细胞外区室均可检测到ROS和RNS。在非疲劳性和疲劳性肌肉收缩过程中其产生均增加。在无疾病的情况下，在骨骼肌中检测到的单个分子种类包括ROS和RNS级联反应的母体自由基：超氧阴离子和一氧化氮。两者的产生速率估计在pmol至nmol/毫克肌肉/分钟范围内。有证据表明，在生理条件下也存在过氧化氢、羟自由基和过氧亚硝酸盐。然而，介导特定生物学效应的分子种类以及肌纤维内ROS和RNS的来源在很大程度上仍未确定。最终明确ROS和RNS调节细胞功能的机制将取决于我们对骨骼肌内ROS和RNS的产生和分布的理解。
