Analysis of glutathione and vitamin C effects on the benzenetriol-induced DNA damage in isolated human lymphocytes.
The alkaline single-cell gel electrophoresis (or Comet) assay was applied to evaluate the eventual DNA damage induced by the triphenolic metabolite of benzene, 1,2,4-benzenetriol (BT), in isolated human lymphocytes. Prior to BT treatment, ranging from 5 to 50 microM, a supplementation with glutathione (GSH, 350 microg/ml) was carried out to assess whether GSH may have a modulating effect on the Comet response. The effect of a fixed dose of BT was also evaluated in the presence of the exogenous antioxidant vitamin C (40 and 200 microM). Additionally, we investigated whether the polymorphism of glutathione S-transferase T1 (GSTT1) gene may affect the individual level of BT-induced DNA damage in vitro. For all donors included in the present study, BT produced a significant dose-response relationship. No clear effect of GSH preincubation was seen on the BT-induced response. On the contrary, a significant reduction of DNA damage was observed in the presence of vitamin C (at least at 200 microM). Although our data suggest some individual differences according to the GSTT1 genotype in the outcome of the Comet assay, a large number of individuals should be studied in further investigations to obtain reliable conclusions.
Pitarque
M
M
Grup de Mutagènesi, Departament de Genètica i de Microbiologia, Edifici Cn, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Creus
A
A
Marcos
R
R
D000328:Adult; D001205:Ascorbic Acid / Q000494:pharmacology*; D001554:Benzene / Q000378:metabolism / Q000633:toxicity*; D002469:Cell Separation; D020552:Comet Assay / Q000379:methods; D004249:DNA Damage / Q000187:drug effects* / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D005978:Glutathione / Q000494:pharmacology*; D006801:Humans; D008214:Lymphocytes / Q000187:drug effects* / Q000502:physiology; D008297:Male
D005978:Glutathione; D001554:Benzene; D001205:Ascorbic Acid