Zinc protects HeLa-tat cells against free radical cytotoxicity induced by glucose.
In the present study, we investigated the protective effect of zinc on the glucose-induced cytotoxicity in HeLa wild and HeLa-tat cells (30 and 20 mmol/l glucose, respectively). HeLa cells transfected with the protein Tat exhibit a lower antioxidant defense system. Incubation of HeLa wild and HeLa-tat cells with high glucose levels led to a rapid increase in generation of reactive oxygen species (ROS). As expected in the presence of high glucose concentrations, the viability was reduced for both cell lines. The redox status essentially regulated by thiol groups may play an important role in the apoptotic process. Thus, we developed a new method using the p-nitrophenyl disulfide to measure cytosolic thiol groups in intact cells. Cellular zinc was measured using inductively coupled plasma mass spectrometry. Intracellular thiol groups and intracellular zinc concentrations were significantly lower in HeLa cells cultured in hyperglycemic conditions, and their concentrations were significantly lower in HeLa-tat cells than in HeLa wild cells. However, the generation of ROS and the induction of apoptosis by a glucose specific mechanism were prevented by zinc (50 micromol/l) and the intracellular thiol groups and zinc concentrations significantly increased in both cell lines to become similar to the initial values. These results suggest that the glucose oxidation and its subsequent effects on the cells can be prevented by a biological antioxidant such as zinc.
Faure
Patrice
P
Laboratoire HP2, Faculté de Médecine, Université Joseph Fourier, 38000 Grenoble, France. patrice.faure@ujf-grenoble.fr
Bouvard
Sophie
S
Roucard
Corinne
C
Favier
Alain
A
Halimi
Serge
S
D002470:Cell Survival / Q000187:drug effects*; D005434:Flow Cytometry; D005609:Free Radicals / Q000037:antagonists & inhibitors*; D015696:Gene Products, tat / Q000378:metabolism*; D005947:Glucose / Q000037:antagonists & inhibitors*; D006367:HeLa Cells; D006801:Humans; D017382:Reactive Oxygen Species / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000378:metabolism; D014162:Transfection; D015032:Zinc / Q000494:pharmacology*
D005609:Free Radicals; D015696:Gene Products, tat; D017382:Reactive Oxygen Species; D013438:Sulfhydryl Compounds; D005947:Glucose; D015032:Zinc