Gomes Ana, Fernandes Eduarda, Lima José L F C
REQUIMTE, Departamento de Química-Física, Faculdade de Farmácia, Universidade do Porto, Rua Aníbal Cunha, 164, 4099-030 Porto, Portugal.
J Biochem Biophys Methods. 2005 Dec 31;65(2-3):45-80. doi: 10.1016/j.jbbm.2005.10.003. Epub 2005 Nov 4.
Endogenously produced pro-oxidant reactive species are essential to life, being involved in several biological functions. However, when overproduced (e.g. due to exogenous stimulation), or when the levels of antioxidants become severely depleted, these reactive species become highly harmful, causing oxidative stress through the oxidation of biomolecules, leading to cellular damage that may become irreversible and cause cell death. The scientific research in the field of reactive oxygen species (ROS) associated biological functions and/or deleterious effects is continuously requiring new sensitive and specific tools in order to enable a deeper insight on its action mechanisms. However, reactive species present some characteristics that make them difficult to detect, namely their very short lifetime and the variety of antioxidants existing in vivo, capable of capturing these reactive species. It is, therefore, essential to develop methodologies capable of overcoming this type of obstacles. Fluorescent probes are excellent sensors of ROS due to their high sensitivity, simplicity in data collection, and high spatial resolution in microscopic imaging techniques. Hence, the main goal of the present paper is to review the fluorescence methodologies that have been used for detecting ROS in biological and non-biological media.
内源性产生的促氧化反应性物种对生命至关重要,参与多种生物学功能。然而,当产生过多(例如由于外源性刺激)或抗氧化剂水平严重耗尽时,这些反应性物种会变得极具危害性,通过生物分子的氧化导致氧化应激,进而造成可能不可逆的细胞损伤并导致细胞死亡。活性氧(ROS)相关生物学功能和/或有害效应领域的科学研究持续需要新的灵敏且特异的工具,以便能更深入地洞察其作用机制。然而,反应性物种具有一些使其难以检测的特性,即它们的寿命非常短,且体内存在多种能够捕获这些反应性物种的抗氧化剂。因此,开发能够克服这类障碍的方法至关重要。荧光探针因其高灵敏度、数据收集简便以及在显微成像技术中的高空间分辨率,是检测ROS的优秀传感器。因此,本文的主要目的是综述已用于在生物和非生物介质中检测ROS的荧光方法。