TY - JOUR
T1 - Effects of vitamins E, C and catalase on bromobenzene- and hydrogen peroxide-induced intracellular oxidation and DNA single-strand breakage in Hep G2 cells
AU - Wu, Jian
AU - Karlsson, Kurt
AU - Danielsson, Åke
PY - 1997/3
Y1 - 1997/3
N2 - Background/aims: Water-soluble vitamin E (Trolox C), ascorbic acid and catalase were shown in our previous study to protect isolated rat hepatocytes against bromobenzene-induced toxicity. Methods: In order to study the mechanisms of this protection and the pathogenesis of bromobenzene-induced hepatocellular injury, a fluorometric assay for the investigation of intracellular oxidation, indicated by conversion of dichlorofluorescein diacetate to dichlorofluorescein, was used. Single-strand DNA breakage was also evaluated in Hep G2 cells by a radio-labelling method. Results: Bromobenzene (2.4 and 4.8 mM) induced a significant increase in dichlorofluorescein fluorescence intensity compared to the controls. Trolox C, ascorbic acid or catalase significantly inhibited bromobenzene-induced enhancement of fluorescence intensity (p < 0.05-0.001), as well as reduced auto-intracellular oxidation in untreated Hep G2 cells. Hydrogen peroxide (H2O2) evoked a dose-dependent increase in dichlorofluorescein fluorescence intensity in Hep G2 cells, and the effect was completely blocked by Trolox C (2.0 mM) and catalase (4800 unit/ml). Bromobenzene caused significant single-strand DNA breakage in Hep G2 cells during 2 h suspension incubation and 24 h primary incubation. H2O2 (400 μM) led to marked single-strand DNA breakage in 20 min, and the effect was attenuated by Trolox C. Conclusions: Metabolism of bromobenzene in Hep G2 cells induces production of H2O2, indicated by enhancement of dichlorofluorescein fluorescence intensity, or other free radicals, which leads to single-strand DNA breakage in the cells. Vitamins E and C and catalase display strong intracellular antioxidative effects. Vitamin E could partially inhibit H2O2-induced single-strand DNA breakage in the cells.
AB - Background/aims: Water-soluble vitamin E (Trolox C), ascorbic acid and catalase were shown in our previous study to protect isolated rat hepatocytes against bromobenzene-induced toxicity. Methods: In order to study the mechanisms of this protection and the pathogenesis of bromobenzene-induced hepatocellular injury, a fluorometric assay for the investigation of intracellular oxidation, indicated by conversion of dichlorofluorescein diacetate to dichlorofluorescein, was used. Single-strand DNA breakage was also evaluated in Hep G2 cells by a radio-labelling method. Results: Bromobenzene (2.4 and 4.8 mM) induced a significant increase in dichlorofluorescein fluorescence intensity compared to the controls. Trolox C, ascorbic acid or catalase significantly inhibited bromobenzene-induced enhancement of fluorescence intensity (p < 0.05-0.001), as well as reduced auto-intracellular oxidation in untreated Hep G2 cells. Hydrogen peroxide (H2O2) evoked a dose-dependent increase in dichlorofluorescein fluorescence intensity in Hep G2 cells, and the effect was completely blocked by Trolox C (2.0 mM) and catalase (4800 unit/ml). Bromobenzene caused significant single-strand DNA breakage in Hep G2 cells during 2 h suspension incubation and 24 h primary incubation. H2O2 (400 μM) led to marked single-strand DNA breakage in 20 min, and the effect was attenuated by Trolox C. Conclusions: Metabolism of bromobenzene in Hep G2 cells induces production of H2O2, indicated by enhancement of dichlorofluorescein fluorescence intensity, or other free radicals, which leads to single-strand DNA breakage in the cells. Vitamins E and C and catalase display strong intracellular antioxidative effects. Vitamin E could partially inhibit H2O2-induced single-strand DNA breakage in the cells.
KW - bromobenzene
KW - catalase
KW - dichlorofluorescein
KW - Hep G cells
KW - hydrogen peroxide
KW - single-strand DNA breakage
KW - vitamin C
KW - vitamin E
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U2 - 10.1016/S0168-8278(97)80434-5
DO - 10.1016/S0168-8278(97)80434-5
M3 - Article
C2 - 9075676
AN - SCOPUS:0031047499
VL - 26
SP - 669
EP - 677
JO - Journal of Hepatology
JF - Journal of Hepatology
SN - 0168-8278
IS - 3
ER -