![Sample our Physical Sciences journals, sign in here to start your FREE access for 14 days]({"type":"image" , "src" : "/sda/110819/TOC-Subject-Sample-2021-Physical-Sciences.jpg"})
Abstract
The fluorogenic probe 2′,7′-dichlorodihydrofluorescein diacetate (H2DCF-DA) is widely used for the estimation of oxidative stress in cells. It is known that 2′,7′-dichlorodihydrofluorescein (H2DCF), product of intracellular hydrolysis of H2DCF-DA, is oxidized to the fluorescent compound, DCF, mainly by hydrogen peroxide (H2O2) in the presence of catalysts. The present study was aimed at answering the question whether the labile iron pool (LIP) may contribute to the oxidation of H2DCF in cellular systems. The membrane-permeable lipophilic iron chelator salicylaldehyde isonicotinoyl hydrazone (SIH) was found to inhibit oxidation of the probe by H2O2 dependent on ferrous ions but not by peroxidase or superoxide dismutase in defined in vitro systems. When applied to cells, the probe inhibited considerably oxidation of H2DCF in V79 Chinese hamster fibroblasts and two murine lymphoma L5178Y(LY) sublines (LY-R, LY-S) differing in LIP level, the extent of inhibition being greater in the LY-R line of higher LIP level. These results demonstrate that LIP is a significant factor determining the rate of intracellular H2DCF oxidation.
| Abbreviations | ||
| DMSO | = | dimethyl sulfoxide |
| HBSS | = | Hank's buffered salt solution |
| H2DCF-DA | = | 2′,7′-dichlorodihydrofluorescein diacetate |
| HPR | = | horseradish peroxidase |
| LIP | = | labile iron pool |
| ROS | = | reactive oxygen species |
| SIH | = | salicylaldehyde isonicotinoyl hydrazone |
| SOD | = | superoxide dismutase |
| Abbreviations | ||
| DMSO | = | dimethyl sulfoxide |
| HBSS | = | Hank's buffered salt solution |
| H2DCF-DA | = | 2′,7′-dichlorodihydrofluorescein diacetate |
| HPR | = | horseradish peroxidase |
| LIP | = | labile iron pool |
| ROS | = | reactive oxygen species |
| SIH | = | salicylaldehyde isonicotinoyl hydrazone |
| SOD | = | superoxide dismutase |