Induction of intestinal cytochrome P450 (CYP3A) by rifampicin in beagle dogs.
Both male and female beagle dogs (four dogs/sex) were orally treated with rifampicin (Rif) at the dose of 10 mg/kg/day for 7 days and an additional eight dogs (four dogs/sex) were used as a control. The inducible effect of Rif on intestinal cytochrome P450, especially CYP3A enzyme, was investigated by measuring microsomal testosterone 6beta-hydroxylation (6beta-OHT) activity, immunoblot and ELISA analysis. In male dogs, microsomal 6beta-OHT activity in the duodenum, upper, middle and lower part of the jejunum and the ileum of the control was 229, 204, 194, 129 and 57 pmol/min/mg protein, while the activity of the Rif-treated dogs significantly increased to 456, 486, 430, 192 and 138 pmol/min/mg protein, respectively. The activity of intestinal 6beta-OHT in the control and Rif-treated female dogs showed almost similar levels to those observed in the corresponding male dogs. The activity of intestinal 6beta-OHT in both control and Rif-treated dogs was specifically inhibited by anti-CYP3A12 antiserum. The apparent K(m) value for 6beta-OHT activity in all sections of the small intestine was comparable with that in the liver, and no significant changes were observed in between control and Rif-treated dogs. In both control and Rif-treated dogs, immunoblotting of intestinal microsomes with anti-CYP3A12 antiserum produced a band indistinguishable from that of purified CYP3A12 or of immunoreactive CYP3A12 in liver microsomes. A significant increase in intestinal CYP3A content by Rif treatment was quantitatively verified by the ELISA analysis and the magnitude of its increase correlated well with that of 6beta-OHT activity elevation. Furthermore, the results of immunohistochemistry using the anti-CYP3A12 antiserum indicated that CYP3A protein was specifically distributed in epithelial cells throughout the small intestine and appeared to be predominant at the apical side of villus cells. These results demonstrate that Rif induces not only hepatic CYP3A12 but also intestinal CYP3A in dogs.
Kyokawa
Y
Y
Developmental Research Laboratories, Shionogi & Co. Ltd., 3-1-1 Futaba-cho, Toyonaka, 561-0825, Osaka, Japan
Nishibe
Y
Y
Wakabayashi
M
M
Harauchi
T
T
Maruyama
T
T
Baba
T
T
Ohno
K
K
D000284:Administration, Oral; D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000276:immunology; D004285:Dogs; D004790:Enzyme Induction; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007106:Immune Sera / Q000494:pharmacology; D015151:Immunoblotting; D007421:Intestine, Small / Q000187:drug effects* / Q000201:enzymology*; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology; D012293:Rifampin / Q000008:administration & dosage / Q000494:pharmacology*; D013250:Steroid Hydroxylases / Q000096:biosynthesis* / Q000276:immunology
D007106:Immune Sera; D003577:Cytochrome P-450 Enzyme System; D013250:Steroid Hydroxylases; C475779:cytochrome P-450 CYP3A12 (canine); C022657:steroid hormone 6-beta-hydroxylase; D012293:Rifampin