Peroxidation-dependent and peroxidation-independent mechanisms by which acetaminophen kills cultured rat hepatocytes.
Acetaminophen killed cultured hepatocytes prepared from male rats induced with 3-methylcholanthrene by two distinct mechanisms. With 0.5 to 5 mM acetaminophen, cell killing within 4 h depended on the inhibition of glutathione reductase by 1,3-bis(chloroethyl)-1-nitrosourea (BCNU) and was accompanied by the peroxidation of cellular lipids as assessed by the accumulation of malondialdehyde. The antioxidant diphenylphenylenediamine (DPPD) prevented both the peroxidation of lipids and the death of the cells. By contrast, DPPD had no effect on the metabolism of acetaminophen as assessed by the extent of the covalent binding of [3H]acetaminophen; by the rate and extent of the depletion of glutathione; and by the accumulation of acetaminophen metabolites in the culture medium. It is concluded that the peroxidation of the phospholipids of cellular membranes is the mechanism whereby 0.5 to 5 mM acetaminophen lethally injures cultured hepatocytes. With 10-20 mM acetaminophen, cell killing at 4 h still depended on BCNU. However, the amount of malondialdehyde in the cultures progressively decreased in parallel with the decreasing ability of DPPD to protect the cells. With 20 mM acetaminophen, there was no evidence of lipid peroxidation, and DPPD had no protective effect. Thus, a second mechanism of lethal cell injury with 10-20 mM acetaminophen is independent of lipid peroxidation and insensitive to antioxidants.
Farber
J L
JL
Department of Pathology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
Leonard
T B
TB
Kyle
M E
ME
Nakae
D
D
Serroni
A
A
Rogers
S A
SA
D000082:Acetaminophen / Q000633:toxicity*; D000818:Animals; D002330:Carmustine / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D008054:Lipid Peroxides / Q000096:biosynthesis*; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008563:Membrane Lipids / Q000378:metabolism; D008748:Methylcholanthrene; D010655:Phenylenediamines / Q000494:pharmacology; D010743:Phospholipids / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013997:Time Factors
D008054:Lipid Peroxides; D008563:Membrane Lipids; D010655:Phenylenediamines; D010743:Phospholipids; D000082:Acetaminophen; D008748:Methylcholanthrene; C004845:N,N'-diphenyl-4-phenylenediamine; D002330:Carmustine