Antioxidant Reactions of Vitamin E in the Perfused Rat Liver: Product Distribution and Effect of Dietary Vitamin E Supplementation

Abstract

We have investigated the relationship between vitamin E (α-tocopherol, TH) oxidation and antioxidant protection in a perfused rat liver model. Perfusion of a male Sprague–Dawley rat liver with 2 m m tert-butylhydroperoxide ( t-BuOOH) for 10 min resulted in lipid peroxidation and metabolic changes reflecting oxidative stress. Mitochondria isolated from the liver exhibited increases in state 3 and state 4 respiration and a decline in the respiratory control ratio. In livers from rats given supplementary vitamin E in the diet, TH content was 7- to 10-fold higher than in controls and lipid peroxidation and metabolic changes induced by t-BuOOH were decreased. In mitochondria from these vitamin E-supplemented livers, the t-BuOOH-induced increase in state 4 respiration was reduced and the respiratory control ratio was maintained. In livers from unsupplemented rats, t-BuOOH induced oxidation of TH to α-tocopherolquinone, α-tocopherolhydroquinone, 2,3-epoxy-α-tocopherolquinone, and 5,6epoxy-α-tocopherolquinone as determined by gas chromatography–mass spectrometry analysis. Yields of these products were approximately doubled by treatment of samples with dilute acid, which indicated the presence of tocopherone and epoxytocopherone precursors. Oxidation of TH in vitamin E-supplemented livers yielded the same products and the relative extent of TH oxidation appeared similar to that in unsupplemented livers. In livers from both unsupplemented and vitamin E-supplemented animals, the distribution of oxidation products was similar in whole liver and isolated mitochondria. These data provide the first simultaneous documentation of TH antioxidant reactions and antioxidant effects in an intact organ system during oxidative stress.