N-acetyl-p-benzoquinone imine-induced changes in the energy metabolism in hepatocytes.
The effect of N-acetyl-p-benzoquinone imine (NAPQI), a reactive metabolite of acetaminophen, on the energy metabolism in isolated hepatocytes was investigated. Incubation of cells with NAPQI (400 microM) resulted in an immediate uptake into the mitochondria, followed by both reduction and glutathione conjugation of the quinone imine. These reactions were extremely rapid and were associated with depletion of the mitochondrial ATP content (greater than 80% depletion after 1 min exposure). The loss of ATP was accompanied by increases in ADP and AMP, as well as NADP. No effect on mitochondrial NAD was observed during this initial phase. Similar alterations were produced by NAPQI in the cytosolic compartment. Furthermore, incubation of hepatocytes with NAPQI inhibited oxygen consumption by nearly 90% within 10 s. In parallel to these biochemical changes, there was marked bleb formation on the surface of the hepatocytes, which was found to precede cell death (trypan blue uptake). In conclusion, our results demonstrate that during exposure of hepatocytes to NAPQI, dramatic changes in cellular energy metabolism occur. These biochemical alterations may be caused by a rapid decrease in mitochondrial function, and they may play an important role in the initiation of NAPQI-induced cytotoxicity.
Andersson
B S
BS
Department of Toxicology, Karolinska Institute, Stockholm, Sweden.
Rundgren
M
M
Nelson
S D
SD
Harder
S
S
D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D016227:Benzoquinones; D004229:Dithiothreitol / Q000494:pharmacology; D004734:Energy Metabolism / Q000187:drug effects*; D007097:Imines / Q000633:toxicity*; D066298:In Vitro Techniques; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D009243:NAD / Q000378:metabolism; D010101:Oxygen Consumption / Q000187:drug effects; D011809:Quinones / Q000633:toxicity*; D051381:Rats; D011919:Rats, Inbred Strains
D016227:Benzoquinones; D007097:Imines; D011809:Quinones; D009243:NAD; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; C028473:N-acetyl-4-benzoquinoneimine; D004229:Dithiothreitol