Noninvolvement of CYP2E1 in the (omega-1)-hydroxylation of fatty acids in rat kidney microsomes.
Pyrazole, acetone, and ethanol are known to induce cytochrome P450 2E1 (CYP2E1) and fatty acid (omega-1)-hydroxylation in rat liver microsomes. However, the nature of the P450 enzyme involved in this (omega-1)-hydroxylation has not been clearly established in extrahepatic tissues such as kidney. Four enzymatic activities (hydroxylations of chlorzoxazone, 4-nitrophenol, and two fatty acids) were assayed in kidney microsomal preparations of rats treated with CYP2E1 inducers. Per os treatment resulted in large increases (threefold to fivefold) in the chlorzoxazone and 4-nitrophenol hydroxylations, and up to a ninefold increase when ethanol was administered by inhalation. However, neither the omega-hydroxylation nor the (omega-1)-hydroxylation of fatty acids was modified. Immunoinhibition specific to CYP2E1 did not significantly decrease the omega and (omega-1)-lauric acid hydroxylations, while the polyclonal anti-CYP4A1 antibody inhibited in part both the omega- and (omega-1)-hydroxylations. Chemical inhibitions using either CYP2E1 competitive inhibitors (such as chlorzoxazone, DMSO, and ethanol) or P450 mechanism-based inhibitors (such as diethyldithiocarbamate and 17-octadecynoic acid) led to a partial inhibition of the hydroxylations. All these results suggest that fatty acid (omega-1)-hydroxylation, a highly specific probe for CYP2E1 in rat and human liver microsomes, is not mediated by CYP2E1 in rat kidney microsomes. In contrast to liver, where two different P450 enzymes are involved in fatty acid omega- and (omega-1)-hydroxylations, the same P450 enzyme, mainly a member of the CYP4A family, was involved in both hydroxylations in rat renal microsomes.
Amet
Y
Y
Laboratoire de Biochimie-Nutrition, Faculté de Médecine, Brest, France. Yolande.Amet-LeBloas@univ-brest.fr
Zerilli
A
A
Goasduff
T
T
Dréano
Y
Y
Berthou
F
F
D000818:Animals; D002753:Chlorzoxazone / Q000378:metabolism; D019392:Cytochrome P-450 CYP2E1 / Q000276:immunology / Q000378:metabolism*; D065691:Cytochrome P-450 CYP2E1 Inhibitors; D042926:Cytochrome P-450 CYP4A; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000276:immunology / Q000494:pharmacology; D005227:Fatty Acids / Q000378:metabolism*; D007158:Immunologic Techniques; D007668:Kidney / Q000201:enzymology*; D008297:Male; D008861:Microsomes / Q000201:enzymology; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D009596:Nitrophenols / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D065691:Cytochrome P-450 CYP2E1 Inhibitors; D004791:Enzyme Inhibitors; D005227:Fatty Acids; D009596:Nitrophenols; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D019392:Cytochrome P-450 CYP2E1; D042926:Cytochrome P-450 CYP4A; D002753:Chlorzoxazone; C024836:4-nitrophenol