Role of vitamin D receptor in the lithocholic acid-mediated CYP3A induction in vitro and in vivo.
Lipophilic bile acids are suggested to be involved in the endogenous expression of CYP3A4 in human and experimental animals as ligands of nuclear receptors. To verify the nuclear receptor specificity, the bile acid-mediated induction of CYP3A4 has been studied in vitro and in vivo in the present study. Lithocholic acid (LCA) strongly enhanced the activities of the CYP3A4 reporter gene, which contained multiple nuclear receptor binding elements, in both HepG2 and LS174T cells. The introduction of small interfering RNA for human vitamin D receptor (VDR), but not for human pregnane X receptor, reduced the LCA-induced activation of the reporter gene in these cells, suggesting the major role of VDR in the LCA induction of CYP3A4. Consistently, oral administration of LCA (100 mg/kg/day for 3 days) increased Cyp3a protein levels in the intestine but not in the liver, where a negligible level of VDR mRNA is detected. The selective role of VDR was tested in mice with the adenoviral overexpression of the receptor. Oral administration of LCA had no clear influence on the CYP3A4 reporter activity in the liver of control mice. In mice with the adenovirally expressed VDR, LCA treatment (100 or 400 mg/kg/day for 3 days) resulted in the enhanced reporter activities and increased levels of Cyp3a proteins in the liver. These results indicate the selective involvement of VDR, but not pregnane X receptor, in the LCA-mediated induction of both human and mouse CYP3As in vivo.
Matsubara
Tsutomu
T
Division of Drug Metabolism and Molecular Toxicology, Graduate School of Pharmaceutical Sciences, Tohoku University, 6-3 Aramaki-aoba, Aoba-ku, Sendai 980-8578, Japan.
Yoshinari
Kouichi
K
Aoyama
Kazunobu
K
Sugawara
Mika
M
Sekiya
Yuji
Y
Nagata
Kiyoshi
K
Yamazoe
Yasushi
Y
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D051544:Cytochrome P-450 CYP3A / Q000096:biosynthesis* / Q000235:genetics; D017931:DNA Primers; D006801:Humans; D008095:Lithocholic Acid / Q000494:pharmacology*; D008099:Liver / Q000187:drug effects / Q000201:enzymology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering; D018167:Receptors, Calcitriol / Q000235:genetics / Q000502:physiology*
D017931:DNA Primers; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D018167:Receptors, Calcitriol; D008095:Lithocholic Acid; D051544:Cytochrome P-450 CYP3A