Allyl alcohol cytotoxicity in isolated rat hepatocytes: mechanism of cell death does not involve an early rise in cytosolic free calcium.
We examined the effect of a toxic concentration of allyl alcohol (0.5 mM) on intracellular calcium concentrations in isolated rat hepatocytes. An increase in phosphorylase a activity was evident in the hepatocytes after 30 min of incubation with allyl alcohol, suggesting that the toxicant may produce an early rise in cytosolic free calcium. The increase in phosphorylase a activity was not reversed by the addition of dithiothreitol (DTT), a sulfhydryl compound that reverses the events that initiate cell killing by allyl alcohol. When intracellular calcium concentrations were measured directly, using fura-2 as the calcium indicator, there was no effect of allyl alcohol on cytosolic free calcium during the first 60 min of exposure, a critical period for development of irreversible damage. Incubation with allyl alcohol did not interfere with the measurement of intracellular calcium. The increases in cytosolic free calcium produced by phenylephrine or ATP were similar to those reported by others and not affected by the presence of allyl alcohol. The results from this study demonstrate that increased cytosolic free calcium is not essential for allyl alcohol-induced cytotoxicity to isolated rat hepatocytes.
Rikans
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LE
Department of Pharmacology, College of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City 73190.
Cai
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Y
Hornbrook
K R
KR
D000433:1-Propanol / Q000633:toxicity; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000704:Analysis of Variance; D000818:Animals; D002118:Calcium / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002712:Chlorides / Q000494:pharmacology; D003600:Cytosol / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D008297:Male; D017895:Manganese Compounds / Q000494:pharmacology; D010656:Phenylephrine / Q000494:pharmacology; D010762:Phosphorylase a / Q000378:metabolism; D020005:Propanols; D051381:Rats; D011916:Rats, Inbred F344
D002712:Chlorides; D017895:Manganese Compounds; D020005:Propanols; D010656:Phenylephrine; C006463:allyl alcohol; D000255:Adenosine Triphosphate; D000433:1-Propanol; D007770:L-Lactate Dehydrogenase; D010762:Phosphorylase a; C025340:manganese chloride; D002118:Calcium; D004229:Dithiothreitol