The effect of dietary selenium and vitamin E on glutathione peroxidase and glutathione in the rat.

نویسندگان

  • D L Scott
  • J Kelleher
  • M S Losowsky
چکیده

There is undoubtedly a close relationship between dietary vitamin E and selenium, but the nature of this is not completely understood. Rotruck et al. (1973) showed that the enzyme glutathione peroxidase (EC 1.1 1.1.9) contains selenium (as an essential cofactor), establishing one of its metabolic roles. Later work established the dependence of tissue glutathione peroxidase activity on dietary selenium intake (Hafeman et al., 1974). The present study investigates the relationships between dietary selenium and vitamin E intakes and tissue glutathione peroxidase activity and glutathione concentration in the rat. Groups of six to eight female Wistar rats were given a basal diet (Kelleher et al., 1972), sufficient in all nutrients and minerals except vitamin E and selenium, from weaning. Separate groups of rats were supplemented with DL-a-tocopherol at 100 and 500mg/kg of diet and/or selenium as sodium selenite at 0.4 and 4p.p.m. After 3 months on these diets, the rats were killed by cardiac puncture under ether anaesthesia, and livers and kidneys removed. Portions of these tissues were homogenized in a glass homogenizer with Teflon pestle, in 2&40vol. of 0.9 % NaCl in O.O~M-EDTA. The homogenates were centrifuged at 750g for 15min at 4"C, and the supernatants then centrifuged at l00OOOg for 60min at 4°C; the soluble fractions were then removed for enzyme assay. The plasma fraction of the blood was removed by centrifugation for enzyme assay, and the erythrocytes were washed three times with an equal volume of 0.9% NaCl in O.O~M-EDTA. Glutathione peroxidase activity was assayed by a modification of the method of Paglia & Valentine (1967) on the soluble tissue fractions and the washed erythrocyte suspensions. GSH (reduced glutathione) was assayed by the method of Sedlak & Lindsay ( I 968) on whole-tissue homogenates and washed erythrocyte suspensions. The protein content of these fractions was assayed by Lowry et a1.k (1951) method, and the erythrocytes were counted on a Coulter counter. Alanine aminotransferase (EC 2.6.1.2) was assayed on the separated plasma.

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عنوان ژورنال:
  • Biochemical Society transactions

دوره 4 2  شماره 

صفحات  -

تاریخ انتشار 1976