25-Hydroxylation of vitamin D3 in primary cultures of pig hepatocytes: evidence for a role of both CYP2D25 and CYP27A1.
There has been some controversy over whether the 25-hydroxylation of vitamin D(3) is carried out by one enzyme or two and whether this cytochrome P450 enzyme is found in the mitochondrial or microsomal fractions of liver. The pig is currently the only species in which both the microsomal 25-hydroxylase (CYP2D25) and the mitochondrial 25-hydroxylase (CYP27A1) have been cloned and characterized. In this paper, the roles of the two enzymes in 25-hydroxylation of vitamin D(3) are examined in primary cultures of hepatocytes. Inhibition experiments indicated that tolterodine and 7 alpha-hydroxy-4-cholesten-3-one were selective inhibitors of the CYP2D25- and CYP27A-mediated 25-hydroxylation of vitamin D(3), respectively. Addition of each inhibitor to primary hepatocytes decreased the total 25-hydroxylation of vitamin D(3) to about the same extent. No inhibition of other hydroxylase activities tested was found. Phorbol 12-myristate 13-acetate down-regulated the expression of both CYP2D25 and CYP27A1 as well as the 25-hydroxylase activity of the hepatocytes. The results implicate that both CYP2D25 and CYP27A1 contribute to the 25-hydroxylation in hepatocytes and are important in the bioactivation of vitamin D(3).
Hosseinpour
Fardin
F
Division of Biochemistry, Department of Pharmaceutical Biosciences, University of Uppsala, Box 578, SE-751 23 Uppsala, Sweden.
Ibranovic
Ines
I
Tang
Wanjin
W
Wikvall
Kjell
K
D000818:Animals; D001559:Benzhydryl Compounds / Q000494:pharmacology; D002478:Cells, Cultured; D002762:Cholecalciferol / Q000378:metabolism*; D053493:Cholestanetriol 26-Monooxygenase; D002783:Cholestenones / Q000494:pharmacology; D003408:Cresols / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism*; D006900:Hydroxylation; D008862:Microsomes, Liver / Q000201:enzymology; D008930:Mitochondria, Liver / Q000201:enzymology; D010665:Phenylpropanolamine; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013250:Steroid Hydroxylases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D034421:Sus scrofa; D013739:Testosterone / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D000068737:Tolterodine Tartrate
D001559:Benzhydryl Compounds; D002783:Cholestenones; D003408:Cresols; D004791:Enzyme Inhibitors; D012333:RNA, Messenger; D002762:Cholecalciferol; D010665:Phenylpropanolamine; C002656:7 alpha-hydroxy-4-cholesten-3-one; D013739:Testosterone; D000068737:Tolterodine Tartrate; D013250:Steroid Hydroxylases; D053493:Cholestanetriol 26-Monooxygenase; D013755:Tetradecanoylphorbol Acetate