Phototoxic lysis of erythrocytes from humans is reduced after oral intake of ascorbic acid and d-α-tocopherol

Ultraviolet (UV) radiation causes hemolysis of human erythrocytes in the presence of photosensitizers. This can be used as an in vitro model for evaluating photosensitizing properties of substances. Antioxidants such as ascorbic acid (vitamin C) and d-alpha-tocopherol (vitamin E) have been found to be photoprotective in such test systems. We assessed the effect of combined systemic intake of both ascorbic acid and d-alpha-tocopherol by human volunteers on phototoxic in vitro lysis of their erythrocytes. In a double-blind placebo-controlled study, 10 subjects took daily 2 g ascorbic acid combined with 1000 IU d-alpha-tocopherol, and 10 took a placebo. Blood was taken before and after 7 days of treatment, erythrocytes were prepared and then incubated with 10(-3) mol/l fenofibrate, a photosensitizer acting in the WA and UVB region. The suspensions were exposed to radiation rich in UVA (up to 40 J/cm(2) UVA) or to radiation rich in UVB (up to 1.6 J/cm(2)). Photohemolysis of the samples was calculated as a percentage of complete hemolysis. At the end of the treatment phase, in the placebo group photohemolysis was not significantly reduced compared with the initial values at all irradiation doses except for 1.6 J/cm(2) UVB (96% vs 79%; P<0.01). In the group taking vitamins, photohemolysis was significantly reduced at nearly all UV doses, most impressively after moderate UVA irradiation (20 J/cm(2) UVA: 86.5% vs 14.5%; P<0.01). It is concluded that the results of the photohemolysis test are influenced by the antioxidative status of the cell donor and that ascorbic acid and d-alpha-tocopherol also may protect against phototoxic damage in vivo.