Glutathione status in primary cultures of rat hepatocytes and its role in cell attachment to collagen.
In primary cultures of rat hepatocytes the intracellular level of reduced glutathione (GSH) declines to approx. 50% of that observed in freshly isolated cells within 1 h of culture. Pretreatment of freshly isolated hepatocytes with diethylmaleate (DEM) to deplete GSH and inhibition of glutathione synthesis by buthionine sulfoximine (BSO) markedly decrease the proportion of cells attaching to the collagen coated culture dishes. A positive correlation between the intracellular content of GSH and the ability of hepatocytes to attach to collagen is observed. Presence of dithiothreitol (DTT) in the culture medium efficiently prevents hepatocyte attachment. A net increase in hepatocyte disulfides is also observed after the first hours of culture. The formation of disulfides seems to be essential for the attachment of hepatocytes to collagen. The depletion of GSH in the early period of culture is probably due to its regulatory function of thiol/disulfide groups in proteins and/or its involvement in the synthesis of essential cytoskeletal proteins.
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D002478:Cells, Cultured; D055598:Chemical Phenomena; D002621:Chemistry; D003094:Collagen / Q000378:metabolism*; D004229:Dithiothreitol / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism / Q000502:physiology*; D007425:Intracellular Membranes / Q000378:metabolism; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008297:Male; D008298:Maleates / Q000494:pharmacology; D010084:Oxidation-Reduction; D051381:Rats; D011919:Rats, Inbred Strains
D008298:Maleates; D003094:Collagen; C030272:maleic acid; D005978:Glutathione; D004229:Dithiothreitol