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Original

Inhibition of Sudan I genotoxicity in human liver-derived HepG2 cells by the antioxidant hydroxytyrosol

, , , , &
Pages 189-195 | Received 20 Oct 2007, Published online: 07 Jul 2009
 

Abstract

The chemoprotective effect of hydroxytyrosol (HT) against Sudan I-induced genotoxicity was investigated in a human hepatoma cell line, HepG2. The comet assay and micronucleus (MN) assay were used to monitor genotoxicity. Intracellular reactive oxygen species (ROS) formation was measured using a fluorescent probe, 2,7-dichlorofluorescein diacetate (DCFH-DA). The levels of oxidative DNA damage and lipid peroxidation were estimated by immunocytochemistry analysis of 8-hydroxydeoxyguanosine (8-OHdG) and by measuring levels of thiobarbituric acid-reactive substances (TBARS), respectively. Intracellular glutathione (GSH) level was estimated by fluorometric methods. The results showed that HT significantly reduced the genotoxicity caused by Sudan I. Furthermore, HT ameliorated lipid pexidation as demonstrated by a reduction in TBARS formation and attenuated GSH depletion in a concentration-dependent manner. It was also found that HT reduced intracellular ROS formation and 8-OHdG level caused by Sudan I. These results strongly suggest that HT has significant protective ability against Sudan I-induced genotoxicity.

Abbreviations
DCFH-DA=

dichlorofluorescein diacetate

EtBr=

ethidium bromide

MN=

micronucleus

8-OHdG=

8-hydroxydeoxyguanosine

HT=

hydroxytyrosol

ROS=

reactive oxygen species

SCGE=

single cell gel electrophoresis

GSH=

glutathione

MDA=

malondialdehyde

TBARS=

thiobarbituric acid-reactive substances

T-BHP=

tert-butylhydroperoxide

LMP=

low melting point

DMSO=

dimethylsulphoxide

NMP=

normal melting point

MEM=

minimum essential Eagle's medium

Abbreviations
DCFH-DA=

dichlorofluorescein diacetate

EtBr=

ethidium bromide

MN=

micronucleus

8-OHdG=

8-hydroxydeoxyguanosine

HT=

hydroxytyrosol

ROS=

reactive oxygen species

SCGE=

single cell gel electrophoresis

GSH=

glutathione

MDA=

malondialdehyde

TBARS=

thiobarbituric acid-reactive substances

T-BHP=

tert-butylhydroperoxide

LMP=

low melting point

DMSO=

dimethylsulphoxide

NMP=

normal melting point

MEM=

minimum essential Eagle's medium

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