Use of a quantitative real-time reverse transcription-polymerase chain reaction method to study the induction of CYPIA, CYP2B and CYP4A forms in precision-cut rat liver slices
article
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 μg ml<sup>-1</sup> Aroclor 1254 (ARO), 500 μM sodium phenobarbitone (NaPB) or 50 μM Wy14,643 (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 24 h 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 24 h 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.
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 μg ml<sup>-1</sup> Aroclor 1254 (ARO), 500 μM sodium phenobarbitone (NaPB) or 50 μM Wy14,643 (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 24 h 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 24 h 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.
Topics
AroclorCytochrome P450 1ACytochrome P450 2BCytochrome P450 4AMessenger RNAPhenobarbitalPirinixic acidXenobiotic agentAnimal tissueControlled studyEnzyme inductionEnzyme regulationLiverMaleNonhumanRatReverse transcription polymerase chain reactionTissue sliceAnimalsAroclorsCytochrome P-450 CYP1A1Cytochrome P-450 CYP1A2Cytochrome P-450 CYP2B1Cytochrome P-450 Enzyme SystemEnvironmental PollutantsExcitatory Amino Acid AntagonistsLiverMaleMutagensPhenobarbitalPyrimidinesRatsRats, Sprague-DawleyReverse Transcriptase Polymerase Chain ReactionRNARNA, MessengerTime FactorsUp-RegulationAnimalia
TNO Identifier
42446
Source
Xenobiotica, 32(9), pp. 739-747.
Pages
739-747
Files
To receive the publication files, please send an e-mail request to TNO Repository.