Increased oxidation of dimethylnitrosamine in pericentral microsomes after pyrazole induction of cytochrome P-4502E1.
The cytochrome P-450IIE1 (CYP2E1) isozyme activates several toxins and procarcinogens. Recent studies employing immunohistochemical and immuno-analysis techniques have shown that this isozyme is predominantly localized in the pericentral zone of the liver acinus. Experiments were conducted to evaluate whether microsomes isolated from the pericentral region of the liver display elevated catalytic activity towards effective substrates for CYP2E1 such as dimethylnitrosamine (DMN) as compared with periportal microsomes. Rats were treated with pyrazole to induce CYP2E1 and hepatocytes prepared from periportal or pericentral zones of the livers by the digitonin-collagenase procedure. Microsomes isolated from these hepatocytes had similar total P-450 contents; however, the microsomes from the pericentral hepatocytes displayed an increased DMSO binding spectrum suggesting an increased content of CYP2E1. Low Km DMN demethylase activity (but not high Km activity) as well as the oxidation of aniline and p-nitrophenol were 2- to 3-fold higher in pericentral compared to periportal microsomes. The oxidation of DMN by both microsomal preparations, as well as the increased rates obtained with the pericentral microsomes, was sensitive to inhibition by carbon monoxide as well as to other CYP2E1 substrates such as ethanol, pyrazole, or 4-methylpyrazole. Anti-CYP2E1 IgG inhibited the oxidation of DMN by both microsomal preparations 75% to 85% and prevented most of the increase found with the pericentral microsomes. Oxidation of aniline and p-nitrophenol was elevated in pericentral hepatocytes compared with periportal hepatocytes to the same extent as in the isolated microsomes.(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Biochemistry, Mount Sinai School of Medicine, New York, New York 10029.
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Dicker
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Cederbaum
D000818:Animals; D046508:Culture Techniques; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis*; D004128:Dimethylnitrosamine / Q000493:pharmacokinetics*; D004790:Enzyme Induction / Q000187:drug effects; D005974:Glutamate-Ammonia Ligase / Q000502:physiology; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects* / Q000201:enzymology*; D011720:Pyrazoles / Q000494:pharmacology*; D051381:Rats; D011919:Rats, Inbred Strains; D005723:gamma-Glutamyltransferase / Q000502:physiology
D011720:Pyrazoles; C031280:pyrazole; D003577:Cytochrome P-450 Enzyme System; D005723:gamma-Glutamyltransferase; D005974:Glutamate-Ammonia Ligase; D004128:Dimethylnitrosamine