Relationships between intracellular vitamin E, lipid peroxidation, and chemical toxicity in hepatocytes.
The cellular content of vitamin E was measured in isolated rat hepatocytes exposed to various types of chemical injury. Vitamin E was determined as alpha-tocopherol by HPLC with in-line uv and electrochemical detection. The cytotoxicity of diquat, a redox cycling compound, was accompanied by a decrease in cellular alpha-tocopherol and a stimulation of lipid peroxidation. Both the loss of alpha-tocopherol and the accumulation of lipid peroxidation products could be prevented by addition of either the antioxidant N,N'-diphenyl-p-phenylenediamine (DPPD) or the reducing agent dithiothreitol (DTT). DTT also prevented the oxidation of soluble and protein thiols and completely protected against cytotoxicity, while DPPD addition only delayed the onset of hepatocyte death. Cytotoxic doses of the naphthoquinone, menadione, and the pyridine compounds 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine and 1-methyl-4-phenyl-pyridinium ion did not deplete alpha-tocopherol levels, nor did they result in significant lipid peroxidation. On the other hand, a peroxidizing, but noncytotoxic dose of ADP-Fe3+ rapidly decreased cellular alpha-tocopherol levels. These data demonstrate that cellular alpha-tocopherol loss is neither a prerequisite for, nor a necessary consequence of toxicity. Moreover, a substantial depletion (ca. 50%) of alpha-tocopherol does not necessarily result in cell death. Although alpha-tocopherol protects against the oxidation of cellular lipids, the maintenance of hepatocyte alpha-tocopherol content does not prevent the oxidation of soluble and protein thiols. These other targets of oxidative damage seem to play a more critical role in hepatocyte toxicity.
Sandy
M S
MS
Department of Biomedical & Environmental Health Sciences, School of Public Health, University of California, Berkeley 94720.
Di Monte
D
D
Smith
M T
MT
D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; D015655:1-Methyl-4-phenylpyridinium; D000244:Adenosine Diphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D002330:Carmustine / Q000494:pharmacology; D004178:Diquat / Q000633:toxicity; D004229:Dithiothreitol / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005978:Glutathione / Q000032:analysis; D066298:In Vitro Techniques; D008054:Lipid Peroxides / Q000378:metabolism*; D008099:Liver / Q000032:analysis / Q000187:drug effects*; D008297:Male; D010655:Phenylenediamines / Q000494:pharmacology; D011725:Pyridines / Q000633:toxicity; D011726:Pyridinium Compounds / Q000633:toxicity; D051381:Rats; D011919:Rats, Inbred Strains; D014810:Vitamin E / Q000032:analysis*
D008054:Lipid Peroxides; D010655:Phenylenediamines; D011725:Pyridines; D011726:Pyridinium Compounds; C029936:adenosine diphosphate-ferric chelate; D014810:Vitamin E; D000244:Adenosine Diphosphate; D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; D004178:Diquat; C004845:N,N'-diphenyl-4-phenylenediamine; D005978:Glutathione; D015655:1-Methyl-4-phenylpyridinium; D004229:Dithiothreitol; D002330:Carmustine