Use of a quantitative real-time reverse transcription-polymerase chain reaction method to study the induction of CYP1A, CYP2B and CYP4A forms in precision-cut rat liver slices.
1. The aim was to employ real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) technology (TaqMan to examine the induction of some selected cytochrome P450 (CYP) forms in precision-cut rat liver slices. 2. Taqman primers and probe sets were developed for rat CYP1A1, CYP1A2, CYP2B1 and CYP4A1 forms. 3. Rat liver slices were cultured in control medium or medium containing either 10 micro g ml(-1) Aroclor 1254 (ARO), 500 micro M sodium phenobarbitone (NaPB) or 50 micro M Wy-14643 (WY) for 3, 6 and 24 h. 4. Compared with control liver slices, treatment with ARO for 3 and 6 h produced 24- and 184-fold increases, respectively, in CYP1A1 mRNA levels, and after 24h produced an 85-fold increase in CYP1A2 mRNA levels. Levels of CYP1A1 and CYP1A2 mRNA were not markedly affected by NaPB and WY. 5. Treatment with ARO and PB for 24 h produced 10.6- and 23.8-fold increases, respectively, in CYP2B1 mRNA. Levels of CYP2B1 mRNA were not markedly affected by WY. 6. Treatment with WY, but not ARO and NaPB, for 24h produced a 20.4-fold increase in levels of CYP4A1 mRNA. 7. These results demonstrate that cultured liver slices may be used to evaluate the effect of xenobiotics on CYP form mRNA levels.
Pan
J
J
Wyeth-Ayerst Research, Drug Metabolism Division, CN8000, Princeton, NJ 08543-8000, USA.
Xiang
Q
Q
Renwick
A B
AB
Price
R J
RJ
Ball
S E
SE
Kao
J
J
Scatina
J A
JA
Lake
B G
BG
D000818:Animals; D001140:Aroclors / Q000494:pharmacology; D019363:Cytochrome P-450 CYP1A1 / Q000096:biosynthesis* / Q000235:genetics; D019388:Cytochrome P-450 CYP1A2 / Q000096:biosynthesis* / Q000235:genetics; D019362:Cytochrome P-450 CYP2B1 / Q000096:biosynthesis* / Q000235:genetics; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000235:genetics; D000072470:Cytochrome P450 Family 4; D004785:Environmental Pollutants; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D008099:Liver / Q000201:enzymology*; D008297:Male; D009153:Mutagens; D010634:Phenobarbital / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D015854:Up-Regulation
D001140:Aroclors; C465387:Cyp4a1protein, rat; D004785:Environmental Pollutants; D018691:Excitatory Amino Acid Antagonists; D009153:Mutagens; D011743:Pyrimidines; D012333:RNA, Messenger; D012313:RNA; C006253:pirinixic acid; D003577:Cytochrome P-450 Enzyme System; D019363:Cytochrome P-450 CYP1A1; D019388:Cytochrome P-450 CYP1A2; D019362:Cytochrome P-450 CYP2B1; D000072470:Cytochrome P450 Family 4; D010634:Phenobarbital