Does the cellular labile iron pool participate in the oxidation of 2′,7′-dichlorodihydrofluorescein?

被引:6
作者
Balcerczyk, Aneta
Kruszewski, Marcin
Bartosz, Grzegorz
机构
[1] Univ Lodz, Dept Mol Biophys, PL-90237 Lodz, Poland
[2] Inst Nucl Chem & Technol, Dept Radiobiol & Hlth Protect, PL-31159 Warsaw, Poland
[3] Maria Sklodowska Curie Mem Canc Ctr, Dept Expt Hematol & Cord Blood Bank, PL-02781 Warsaw, Poland
[4] Inst Oncol, PL-02781 Warsaw, Poland
[5] Univ Rzeszow, Dept Biochem & Cell Biol, PL-35310 Rzeszow, Poland
关键词
2; 7 '-dichlorodihydrofluorescein diacetate; iron chelation; salicylaldehyde isonicotinoyl hydrazone (SIH); labile iron pool;
D O I
10.1080/10715760601175353
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:563 / 570
页数:8
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