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Am J Physiol Gastrointest Liver Physiol 258: G967-G973, 1990;
0193-1857/90 $5.00
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AJP - Gastrointestinal and Liver Physiology, Vol 258, Issue 6 967-G973, Copyright © 1990 by American Physiological Society

ARTICLES

Inhibition of GSH efflux from rat liver by methionine: effects of GSH synthesis in cells and perfused organ

J. C. Fernandez-Checa, T. Maddatu, M. Ookhtens and N. Kaplowitz
Liver Research Laboratory, Wadsworth Veterans Administration Hospital Center, Los Angeles 90073.

The inhibition of efflux of intracellular reduced glutathione (GSH) by methionine was determined in isolated rat hepatocytes suspended either in Krebs-Henseleit buffer or in modified Fisher's medium. Methionine (1 mM) added to Krebs-Henseleit suspensions of isolated rat hepatocytes inhibited GSH efflux, with greater retention of GSH in the cells compared with control. Results were similar with methionine and 0.3 mM propargylglycine cystathionase inhibitor), suggesting no net synthesis of GSH from methionine. In Fisher's medium, the inhibitory effect of methionine on GSH efflux was masked due to increasing cellular GSH; however, the inhibitory effect of methionine was unmasked by propargylglycine, which prevented the utilization of methionine for GSH synthesis. The addition of serine (0.1 mM) to methionine in Krebs-Henseleit buffer raised cellular GSH, overcoming the inhibition of GSH efflux. In the perfused liver, infusion of 1 and 5 mM methionine initially inhibited GSH efflux, but the inhibition was reversed with continued methionine infusion. After removal of methionine, GSH efflux increased immediately. The reversal and rebound were blocked by propargylglycine, revealing concentration-dependent inhibition of sinusoidal GSH efflux by methionine. Thus, when methionine is utilized to promote GSH synthesis, its inhibitory effect on GSH efflux tends to be overcome.


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