Oyama Y, Hayashi A, Ueha T, Maekawa K
Laboratory of Cell Signaling (Pharmacological Science), Faculty of Integrated Arts and Sciences, University of Tokushima, Japan.
Brain Res. 1994 Jan 28;635(1-2):113-7. doi: 10.1016/0006-8993(94)91429-x.
The fluorescence of 2',7'-dichlorofluorescin (DCF) was measured in acutely dissociated rat cerebellar neurons as a mean of estimating the formation of reactive oxygen species (ROS). N,N-Diethyldithiocarbamate, an inhibitor for superoxide dismutase, reduced the intensity of DCF fluorescence in a dose-dependent fashion at concentrations of 30 nM to up to 10 microM. N-Ethylmaleimide, an inhibitor for glutathione peroxidase, augmented the DCF fluorescence in a dose-dependent manner at concentration of 10 microM to 1 mM while 3-amino-1,2,4-triazole, an inhibitor for catalase, did not change the fluorescence intensity even at concentrations as high as 1 mM. Hydrogen peroxide, applied externally at concentrations between 3 microM and 3 mM, augmented the fluorescence in a dose-dependent fashion. These results suggest the possibility that the DCF fluorescence may be useful in estimating the intracellular content of hydrogen peroxide of mammalian brain neurons.
通过测量急性分离的大鼠小脑神经元中2',7'-二氯荧光素（DCF）的荧光，来评估活性氧（ROS）的生成。超氧化物歧化酶抑制剂N,N-二乙基二硫代氨基甲酸盐在30 nM至10 μM的浓度范围内，以剂量依赖的方式降低了DCF荧光强度。谷胱甘肽过氧化物酶抑制剂N-乙基马来酰亚胺在10 μM至1 mM的浓度范围内，以剂量依赖的方式增强了DCF荧光，而过氧化氢酶抑制剂3-氨基-1,2,4-三唑即使在高达1 mM的浓度下也未改变荧光强度。外部施加浓度在3 μM至3 mM之间的过氧化氢，以剂量依赖的方式增强了荧光。这些结果表明，DCF荧光可能有助于评估哺乳动物脑神经元内过氧化氢的含量。
