Effect of dietary vitamin E and selenium on DNA damage in fresh and frozen tissues

To determine whether deficiencies of dietary vitamin E and Se can elevate background DNA damage, rats were fed diets deficient in or supplemented with vitamin E (30 and 200 mg/kg diet) and Se (0.2 mg/kg diet) for 8 weeks. DNA damage was measured using the Comet (single-cell electrophoresis) assay and 8-oxodeoxyguanosine (8-oxo-dG) in liver, kidneys, and lymphocytes. We found that a deficiency of vitamin E and/or Se for 8 weeks did not significantly increase DNA damage in freshly isolated liver, kidneys, or lymphocytes. However, deficiency of vitamin E and/or Se for 8 weeks markedly increased DNA strand breaks in frozen kidney (-80 degrees C for 72 hours) and in lymphocytes incubated overnight at 37 degrees C, both of which were effectively prevented by supplementation of Se and vitamin E. However, vitamin E at 200 mg/kg did not afford more protection than it did at 30 mg/kg). Little or no significant increase in DNA damage was found in frozen livers. These results indicate that freezing or freeze-thawing of tissues may cause oxidative damage to DNA when the tissues are deficient in a major antioxidant, and that normal levels of vitamin E (30 mg/kg diet) and Se (0.2 mg/kg diet) are sufficient to prevent the damage. Thus, our results caution against the interpretation of DNA data obtained from frozen rat tissues or cells in animal studies with dietary vitamin E or Se deficiencies.