Drake Danielle M, Shapiro Aaron M, Wells Peter G
Department of Pharmaceutical Sciences and Centre for Pharmaceutical Oncology, University of Toronto, Toronto, ON, Canada.
Department of Pharmaceutical Sciences, University of Toronto, Toronto, ON, Canada.
Methods Mol Biol. 2019;1965:313-328. doi: 10.1007/978-1-4939-9182-2_21.
Reactive oxygen species (ROS) can oxidize cellular macromolecules like DNA, causing DNA damage. The most common form of DNA damage is the 8-oxoguanine (8-oxoG) lesion, typically repaired by the base excision repair (BER) pathway, which is initiated by the enzyme oxoguanine glycosylase 1 (OGG1). ROS are produced endogenously and can be enhanced by environmental factors, such as xenobiotics, radiation, and microbial pathogens. As a commonly used biomarker of oxidative damage, 8-oxoG can be measured in two different ways described herein. Commercially available ELISA kits allow for easy detection of the 8-oxoG lesion, while more difficult HPLC assays with UV and electrochemical detection allow for a more definitive identification and quantification of 8-oxoG.
活性氧(ROS)可氧化细胞大分子如DNA,导致DNA损伤。DNA损伤最常见的形式是8-氧代鸟嘌呤(8-oxoG)损伤,通常由碱基切除修复(BER)途径修复,该途径由氧代鸟嘌呤糖基化酶1(OGG1)启动。ROS是内源性产生的,并且可被环境因素增强,如外源性物质、辐射和微生物病原体。作为氧化损伤的常用生物标志物,8-oxoG可通过本文所述的两种不同方法进行测量。市售的ELISA试剂盒可轻松检测8-oxoG损伤,而更具难度的采用紫外和电化学检测的高效液相色谱法(HPLC)可对8-oxoG进行更明确的鉴定和定量。