Modulating effect of glutathione disulfide on thyroxine-5'-deiodination by rat hepatocytes in primary culture: effect of glucose.
To investigate whether an increase in the intracellular glutathione disulfide (GSSG) concentration actually regulates T4-5'-deiodination in intact cells, rat hepatocytes in primary culture were exposed to glutathione-oxidizing agents (diamide and tertiary butylhydroperoxide) or vinblastine, and their effects on 5'-deiodination of T4 were studied. Deiodinating activity was determined from the 125I- fraction released from [3',5'-125I]T4 added to the serum-free culture medium. Total glutathione (T-GSH) and GSSG levels were determined enzymatically. Diamide (1 mM) and tertiary butylhydroperoxide (0.5 mM) increased the GSSG fraction to approximately 40% of the T-GSH at 5 min, followed by a rapid decrease in GSSG. Glucose deprivation of the medium caused a greater GSSG level at 5 min, followed by a delayed normalization of the increased GSSG level. T4-5'-deiodinating activity was minimally decreased in hepatocytes exposed to 1 mM diamide in the presence of glucose in the medium, but was significantly inhibited in the absence of glucose. Vinblastine, in contrast, gradually and steadily increased the GSSG fraction, and by 3 h, GSSG exceeded 20% of T-GSH (at 10(-4) M vinblastine). This was accompanied by a significant inhibition of 5'-deiodinating activity. When the enzyme activity was inhibited, the T-GSH level was decreased to 40-80% of the control level, which per se cannot account for the decreased T4-5'-deiodinating activity, as reported previously. These data suggest that the increased GSSG level, but not the T-GSH concentration, modulates T4-5'-deiodination in intact cells, and that glucose stimulates the enzyme activity by maintaining glutathione in the reduced form, probably through supplying NADPH, a cofactor for GSSG reductase.
D000818:Animals; D002478:Cells, Cultured; D003958:Diamide / Q000494:pharmacology; D005947:Glucose / Q000494:pharmacology*; D005978:Glutathione / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D019803:Glutathione Disulfide; D007455:Iodine / Q000378:metabolism*; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D010545:Peroxides / Q000494:pharmacology; D051381:Rats; D013974:Thyroxine / Q000378:metabolism*; D014747:Vinblastine / Q000494:pharmacology; D020122:tert-Butylhydroperoxide
D010545:Peroxides; D003958:Diamide; D014747:Vinblastine; D020122:tert-Butylhydroperoxide; D007455:Iodine; D005978:Glutathione; D005947:Glucose; D013974:Thyroxine; D019803:Glutathione Disulfide