Close Graeme L, Jackson Malcolm J
Division of Metabolic and Cellular Medicine, School of Clinical Sciences, University of Liverpool, Liverpool, UK.
Methods Mol Biol. 2008;477:123-36. doi: 10.1007/978-1-60327-517-0_11.
Reactive oxygen species (ROS) are constantly produced by skeletal muscle and this production is increased during contractile activity. Understanding the role that ROS play in skeletal muscle requires an understanding of the species of ROS produced, the subcellular site of production, the time-course of ROS production, and the effects of inhibiting these ROS using specific antioxidants or inhibitors. Unfortunately, due to the extremely short half-lives of ROS, many methods to study ROS have had to rely on downstream markers of ROS reactions which cannot provide specific information. In vivo microdialysis is one technique that allows access to a specific site of ROS production allowing the continuous measurement of ROS at rest and during contractile activity. This chapter will describe the technique of microdialysis to measure ROS in skeletal muscle as well as discussing specific methods to detect superoxide, nitric oxide, and hydroxyl radical activity using in vivo microdialysis in skeletal muscle at rest and during contractile activity.
活性氧(ROS)由骨骼肌持续产生,且在收缩活动期间这种产生会增加。要了解ROS在骨骼肌中所起的作用,需要了解所产生的ROS种类、产生的亚细胞位点、ROS产生的时间进程,以及使用特定抗氧化剂或抑制剂抑制这些ROS的效果。不幸的是,由于ROS的半衰期极短,许多研究ROS的方法不得不依赖于ROS反应的下游标志物,而这些标志物无法提供具体信息。体内微透析是一种能够进入ROS产生的特定部位的技术,可在静息和收缩活动期间连续测量ROS。本章将描述用于测量骨骼肌中ROS的微透析技术,并讨论在静息和收缩活动期间使用体内微透析检测骨骼肌中超氧化物、一氧化氮和羟自由基活性的具体方法。