Galactosamine-induced cell death in primary cultures of rat hepatocytes.
Primary cultures of rat hepatocytes were exposed to 0.5 mM D-galactosamine. After 36 hours, only 10-20% of the original cells were viable, as assessed by trypan blue exclusion. In the absence of galactosamine, there was no loss of viability over this same period. The addition of 3 mM uridine to the culture medium completely prevented the cell death produced by galactosamine. Glucosamine had no effect on the viability of the hepatocytes. The extent of galactosamine-induced cell death was dependent upon the concentration of Ca++ ions in the culture medium. With the only source of Ca++ that added with the fetal calf serum, galactosamine had only a very slight effect on viability. With higher Ca++ than with the fetal calf serum, galactosamine had only a very slight effect on viability. With higher Ca++ concentrations, from 0.9 to 3.6 mM, the viability ranged from 75% to 31% 18 hours after treatment with galactosamine. The addition of 1.4 microM chlorpromazine to culture medium containing 1.8 mM Ca++ decreased the extent of the galactosamine-induced cell death. This protective effect was progressively reduced by raising the Ca++ concentration to 3.6 and 5.4 mM. Chlorpromazine given to intact rats 2 hours after treatment with 400 mg/kg galactosamine prevented the appearance of liver cell necrosis. At the same time, chlorpromazine prevented the increases in liver cell Ca++ content. These results indicate that many of the features of the effect of galactosamine on intact rat liver cells can be reproduced in primary cultures of these same cells. The data also support the hypothesis that a disturbance in intracellular Ca++ homeostasis leading to accumulations of these ions is causally related to the cell death produced by galactosamine.
D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects*; D002746:Chlorpromazine / Q000494:pharmacology; D046508:Culture Techniques; D005260:Female; D005688:Galactosamine / Q000494:pharmacology*; D007477:Ions; D008099:Liver / Q000166:cytology*; D051381:Rats; D014529:Uridine / Q000494:pharmacology
D007477:Ions; D005688:Galactosamine; D002118:Calcium; D002746:Chlorpromazine; D014529:Uridine