Waterfall A H, Singh G, Fry J R, Marsden C A
Department of Physiology and Pharmacology, Queen's Medical Centre, Nottingham, UK.
Brain Res Brain Res Protoc. 1997 Dec 1;2(1):17-22. doi: 10.1016/s1385-299x(97)00022-6.
Tissue damage due to free radical activity may be a common mediator in pathology [Kehrer, Crit. Rev. Toxicol. 23 (1993) 21-48]. The confirmation that free radicals have a role in the disease process is hampered by the difficulty of their measurement, as free radicals are transient species present at low concentrations in tissue. Direct measurement of free radicals is confined to electron spin resonance, a specialised methodology not available in our laboratory. We have evaluated several indirect measures, namely, changes in antioxidant status, hydroxyl radical trapping assays and monitoring breakdown products of free radical attack. Here we describe a method from the latter category, monitoring lipid peroxidation. The measurement of lipid peroxidation has been favoured as the indicator of free radical activity in previous ex vivo and in vitro studies of oxidative stress, in particular by the measurement of malondialdehyde (MDA). There is an absence of reported measurements of MDA from in vivo preparations especially where samples are collected in situ. The method describes the combination of microdialysis sampling with high performance liquid chromatography coupled to ultraviolet spectroscopy, to measure MDA from in vivo preparations of rat brain. The methodology could equally be applied to other tissues.
自由基活性导致的组织损伤可能是病理学中一种常见的介导因素[凯勒，《毒理学评论》23（1993年）21 - 48页]。由于自由基是在组织中以低浓度存在的瞬态物质，其测量难度使得自由基在疾病过程中作用的确认受到阻碍。自由基的直接测量仅限于电子自旋共振，这是我们实验室无法使用的一种专业方法。我们评估了几种间接测量方法，即抗氧化状态的变化、羟基自由基捕获测定以及监测自由基攻击的分解产物。在此我们描述后一类方法中的一种，即监测脂质过氧化。在先前关于氧化应激的体外和体内研究中，脂质过氧化的测量一直被用作自由基活性的指标，特别是通过测量丙二醛（MDA）。目前尚无关于体内制剂中MDA测量的报道，尤其是在原位采集样本的情况下。该方法描述了微透析采样与高效液相色谱 - 紫外光谱联用，用于测量大鼠脑体内制剂中的MDA。该方法同样可应用于其他组织。
