Eruslanov Evgeniy, Kusmartsev Sergei
Shands Cancer Center and Department of Urology, University of Florida, Gainesville, FL, USA.
Methods Mol Biol. 2010;594:57-72. doi: 10.1007/978-1-60761-411-1_4.
Cells constantly generate reactive oxygen species (ROS) during aerobic metabolism. The ROS generation plays an important protective and functional role in the immune system. The cell is armed with a powerful antioxidant defense system to combat excessive production of ROS. Oxidative stress occurs in cells when the generation of ROS overwhelms the cells' natural antioxidant defenses. ROS and the oxidative damage are thought to play an important role in many human diseases including cancer, atherosclerosis, other neurodegenerative diseases and diabetes. Thus, establishing their precise role requires the ability to measure ROS accurately and the oxidative damage that they cause. There are many methods for measuring free radical production in cells. The most straightforward techniques use cell permeable fluorescent and chemiluminescent probes. 2'-7'-Dichlorodihydrofluorescein diacetate (DCFH-DA) is one of the most widely used techniques for directly measuring the redox state of a cell. It has several advantages over other techniques developed. It is very easy to use, extremely sensitive to changes in the redox state of a cell, inexpensive and can be used to follow changes in ROS over time.
细胞在有氧代谢过程中持续产生活性氧（ROS）。ROS的产生在免疫系统中发挥着重要的保护和功能作用。细胞配备了强大的抗氧化防御系统来对抗ROS的过量产生。当ROS的产生超过细胞的天然抗氧化防御能力时，细胞内就会发生氧化应激。ROS和氧化损伤被认为在包括癌症、动脉粥样硬化、其他神经退行性疾病和糖尿病在内的许多人类疾病中起重要作用。因此，要确定它们的确切作用，就需要有准确测量ROS及其所造成氧化损伤的能力。有许多方法可用于测量细胞中的自由基产生。最直接的技术是使用可透过细胞的荧光和化学发光探针。2'-7'-二氯二氢荧光素二乙酸酯（DCFH-DA）是直接测量细胞氧化还原状态最广泛使用的技术之一。与其他已开发的技术相比，它有几个优点。它使用非常简便，对细胞氧化还原状态的变化极其敏感，成本低廉，并且可用于追踪ROS随时间的变化。
