Open AccessGlutathione Efflux from Perfused Rat Liver after Phenobarbital Treatment, during Drug Oxidations, and in Selenium Deficiency
Author(s) -
SIES Helmut,
BARTOLI Gianna M.,
BURK Raymond F.,
WAYDHAS Christian
Publication year - 1978
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1978.tb20902.x
Subject(s) - glutathione , efflux , chemistry , phenobarbital , glutathione peroxidase , endocrinology , medicine , ethylmorphine , selenium , lipid peroxidation , biochemistry , metabolism , antioxidant , drug metabolism , biology , enzyme , organic chemistry
1 The efflux of reduced (GSH) and oxidized (GSSG) glutathione from perfused rat liver was studied in different metabolic conditions. 2 At 37 °C in non‐recirculating hemoglobin‐free perfusion, efflux of glutathione per gram of liver was 14 nmol/min, of which GSH was 12 nmol/min and GSSG 1 nmol/min. Similar rates were observed in livers from phenobarbital‐pretreated rats. This is interpreted to indicate that the increased capacity for H 2 O 2 production described in the literature for isolated microsomal membranes is not detectably realized in the intact cell, since H 2 O 2 production from other intracellular sources, e.g. benzylamine, leads to an extra GSSG efflux. Similarly, the glutathione release from livers of selenium‐deficient rats low in GSH peroxidase (8% of the selenium‐supplemented controls, H 2 O 2 as substrate) was not significantly different from the controls. This observation contrasts with that of Burk et al. [ J. Biol. Chem. 253 , 43–46] who found a 2‐fold increase in glutathione release in livers from rats exposed to selenium‐deficiency for a more extended time period. Thus, elevated rates of lipid peroxidation may occur in more severe selenium‐deficiency where factors other than a low GSH peroxidase activity may contribute to the effect. 3 An increased rate of GSSG efflux was observed in livers from phenobarbital‐pretreated rats during drug oxidations dependent on cytochrome P ‐450, using aminopyrine, ethylmorphine and hexobarbital as substrates. Half‐maximal aminopyrine concentration for GSSG efflux, 0.1–0.2 mM, was similar to that of the extra O 2 uptake but lower than that for aminopyrine N‐demethylation, 0.5–0.6 mM. Aminopyrine‐dependent GSSG efflux was present also in livers from selenium‐deficient rats whereas H 2 O 2 ‐dependent or benzylamine‐dependent GSSG efflux was abolished. These results do not support extra production of free H 2 O 2 during drug oxidation in the intact cell. Future investigations must evaluate whether glutathione oxidation occurs by reactive drug metabolites or by an enhanced rate of lipid peroxidation, or by back‐equilibration of glutathione reductase concomitant with the decreased NADPH levels.