Metabolism of human cytochrome P450 marker substrates in mouse: a strain and gender comparison.
The aim was to characterize mouse gender and strain differences in the metabolism of commonly used human cytochrome (CYP) P450 probe substrates. Thirteen human CYP probe substrates (phenacetin, coumarin, 7-ethoxy-4-trifluoromethyl coumarin, amiodarone, paclitaxel, diclofenac, S-mephenytoin, bufuralol, dextromethorphan, chlorzoxazone, p-nitrophenol, testosterone and lauric acid) were used in activity measurements. The metabolism of the probe substrates was compared in liver microsomes from male and female NMRI, CBA, C57bl/6, 129/SvJ and CD1 strains. The expression of proteins identified on Western blots with commonly available antibodies selective for specific human and rat CYP enzymes were compared in the different mouse strains. Males had higher metabolism than corresponding females for phenacetin O-deethylation (human marker for CYP1A2 activity), and a high correlation was found between phenacetin activity and immunoreactivity in Western blots produced with rat CYP1A2 antibodies. Protein detected by antibodies cross-reacting with human CYP2B6 and rat CYP2B1/2 antibodies was female specific except for the 129/SvJ strain, where it was absent in both genders. Females generally had a higher metabolism of bufuralol 1'-hydroxylation and dextromethorphan O-demethylation (human markers for CYP2D activity). Bufuralol 1'-hydroxylation correlated with a female-dominant mouse CYP, which was detected with antibodies against rat CYP2D4. p-Nitrophenol 2-hydroxylation correlated better than chlorzoxazone 6-hydroxylation with the protein detected with antibodies against rat CYP2E1, indicating that p-nitrophenol is a more specific substrate for mouse CYP2E1.
Löfgren
S
S
Department of Biomedical Sciences and Veterinary Public Health, Division of Pathology, Pharmacology and Toxicology, Swedish University of Agricultural Sciences, Box 7028, SE-750 07 Uppsala, Sweden. susanne.lofgren@pat.slu.se
Hagbjörk
A L
AL
Ekman
S
S
Fransson-Steen
R
R
Terelius
Y
Y
D000638:Amiodarone / Q000378:metabolism; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000096:biosynthesis; D015153:Blotting, Western; D002753:Chlorzoxazone / Q000378:metabolism; D003374:Coumarins / Q000378:metabolism; D019388:Cytochrome P-450 CYP1A2 / Q000096:biosynthesis; D065702:Cytochrome P-450 CYP2B6; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D018076:DNA, Complementary / Q000378:metabolism; D003915:Dextromethorphan / Q000378:metabolism; D004008:Diclofenac / Q000378:metabolism; D004983:Ethanolamines / Q000378:metabolism; D005260:Female; D006801:Humans; D007700:Kinetics; D007850:Lauric Acids / Q000378:metabolism; D008297:Male; D008617:Mephenytoin / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008862:Microsomes, Liver; D006899:Mixed Function Oxygenases / Q000096:biosynthesis; D009249:NADP / Q000378:metabolism; D009596:Nitrophenols / Q000378:metabolism; D010089:Oxidoreductases, N-Demethylating / Q000096:biosynthesis; D010100:Oxygen / Q000378:metabolism; D017239:Paclitaxel / Q000378:metabolism; D010615:Phenacetin / Q000378:metabolism; D051381:Rats; D012737:Sex Factors; D013045:Species Specificity; D013379:Substrate Specificity; D013739:Testosterone / Q000378:metabolism
D003374:Coumarins; D018076:DNA, Complementary; D004983:Ethanolamines; D007850:Lauric Acids; D009596:Nitrophenols; C056033:7-ethoxy-4-trifluoromethylcoumarin; C030358:lauric acid; D004008:Diclofenac; D013739:Testosterone; D009249:NADP; D003915:Dextromethorphan; C010831:bufuralol; D003577:Cytochrome P-450 Enzyme System; C030123:coumarin; D006899:Mixed Function Oxygenases; D001189:Aryl Hydrocarbon Hydroxylases; C585599:CYP2B6 protein, human; C115670:Cyp2d4 protein, rat; D019388:Cytochrome P-450 CYP1A2; D065702:Cytochrome P-450 CYP2B6; D010089:Oxidoreductases, N-Demethylating; D010615:Phenacetin; D002753:Chlorzoxazone; D000638:Amiodarone; D017239:Paclitaxel; D008617:Mephenytoin; D010100:Oxygen; C024836:4-nitrophenol