Differential effect of cyanohydroxybutene on glutathione synthesis in liver and pancreas of male rats

1-Cyano-2-hydroxy-3-butene (CHB), an aliphatic nitrile found in cruciferous vegetables, causes a two- and sevenfold elevation in reduced glutathione (GSH) in rat liver and pancreas, respectively, after oral administration of 200 mg/kg. While this dose is also associated with pancreatotoxicity, a single 100 mg/kg dose or multiple lesser doses show the same effect, although somewhat reduced in magnitude, with no concomitant toxicity. In an attempt to identify the mechanism of this increase, we investigated the effect of CHB on GSH synthesis by examining the effect of buthionine sulfoximine (BSO), an inhibitor of GSH synthesis, on CHB-induced GSH elevation. Male Fischer 344 rats received 3 mmol BSO/kg ip 24 and 34 hr following CHB or corn oil. The CHB-mediated elevation in hepatic and pancreatic GSH was eradicated by BSO, suggesting that increased synthesis was responsible. The rate-limiting step in synthesis is gamma-glutamyl cysteine synthetase (GCS); the limiting substrate is cysteine. Therefore, CHB effects on GCS activity and hepatic and pancreatic cysteine equivalents were investigated. When rats were treated by gavage with CHB (100 mg/kg), hepatic GCS mRNA concentrations were increased 24 hr after treatment and hepatic cysteine equivalents were significantly elevated 4 hr following CHB. No significant elevation in hepatic GCS activity was observed, however, even 24 hr following CHB. Pancreatic cysteine equivalents were elevated at both 4 and 8 hr after CHB treatment. However, there was no detectable GCS mRNA or activity in pancreas, in either control or treated animals. Furthermore, CHB had no direct effect on the activity of GCS purified from kidney, regardless of whether GSH was present or absent. These results suggest that the mechanism of CHB-mediated induction of GSH may involve early increases in GSH precursors as well as a later increase in GCS mRNA. The mechanism of GSH elevation identified in these studies may hold therapeutic or prophylactic implications.