Cysteamine increases homocysteine export and glutathione content by independent mechanisms in C3H/10T1/2 cells.
Several thiols, including homocysteine and cysteamine, have been shown to increase glutathione levels in C3H/10T1/2 Cl 8 cells [Biochem. Pharmacol. 39:421-429 (1990)]. The present paper shows that cysteamine also increases homocysteine export from these cells. Cellular glutathione content and export of glutathione and homocysteine increased with increasing doses of cysteamine. Twenty-four hours after addition, 300 microM cysteamine increased both glutathione content and homocysteine export 3-4-fold. No change in the ratio between reduced and oxidized glutathione could be detected, suggesting that the cysteamine effect was not due to reduction of pools of oxidized glutathione. The elevation of glutathione occurred rapidly but declined between 24 and 48 hr after addition of cysteamine, whereas the homocysteine export increased momentarily after cysteamine exposure and then proceeded at a rate similar to that from untreated control cells. The cysteamine-induced increase in glutathione was completely prevented by the gamma-glutamylcysteine synthetase inhibitor buthionine sulfoximine but was not affected by inhibition of homocysteine formation by 3-deazaaristeromycin. Buthionine sulfoximine did not prevent the increase in homocysteine export by cysteamine, and only a small increase in homocysteine export was observed when the cells were exposed to 3-deazaaristeromycin before treatment with cysteamine. Two major conclusions were drawn. 1) Increase of glutathione content and homocysteine export by cysteamine were independent events, indicating that glutathione status and homocysteine formation are regulated by independent mechanisms in C3H/10T1/2 Cl 8 cells. 2) S-Adenosylhomocysteine catabolism was the main source of the homocysteine export induced by cysteamine.
Djurhuus
R
R
Department of Pharmacology and Toxicology, University of Bergen, Norway.
Svardal
A M
AM
Ueland
P M
PM
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002478:Cells, Cultured; D003543:Cysteamine / Q000494:pharmacology*; D003545:Cysteine / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005978:Glutathione / Q000378:metabolism*; D006710:Homocysteine / Q000378:metabolism*; D051379:Mice
D006710:Homocysteine; D003543:Cysteamine; D005978:Glutathione; D003545:Cysteine