In the present study, time-dependency of the induction effect of a selective inducer on the activity, protein and mRNA levels of cytochromes P450 1A1/2 (CYP1A1/2), NAD(P)H:quinone oxidoreductase 1 (NQO1) and glutathione S-transferases (GSTA), in primary culture of rat hepatocytes was tested and evaluated. To show the differences in responses of tested enzymes, the common aryl hydrocarbon receptor (AhR) ligand agonist, beta-naphthoflavone (BNF), was used. Induction of CYP1A1/2 by BNF was detected at all time intervals and at all levels (i.e., mRNA, protein, enzyme activity). Different responses of NQO1 and GSTA upon BNF treatment were observed. Our results demonstrate that the responses of different xenobiotic-metabolizing enzymes to the inducer vary in time and depend on the measured parameter. For these reasons, an induction study featuring only one-time interval treatment and/ or one parameter testing could produce misleading information.
If the inline PDF is not rendering correctly, you can download the PDF file here.
1. H. Fujimura N. Murakami S. Miwa C. Aruga and W. Toriumi The suitability of rat hepatoma cell line H4IIE for evaluating the potentials of compounds to induce CYP3A23 expression Exp. Toxicol. Pathol. 64 (2012) 527-533; https://doi.org/10.1016/j.etp.2010.11.010
2. D. B. Buckley and C. D. Klaassen Induction of mouse UDP-glucuronosyltransferase mRNA expression in liver and intestine by activators of aryl-hydrocarbon receptor constitutive androstane receptor pregnane X receptor peroxisome proliferator-activated receptor alpha and nuclear factor erythroid 2-related factor 2 Drug Metab. Dispos. 37 (2009) 847-856; https://doi.org/10.1124/dmd.108.024190
3. Food and Drug Administration Guidance for Industry: Drug Interaction Studies - Study Design Data Analysis and Implications for Dosing and Labeling Draft Guidance FDA Center for Drug Evaluation and Research Rockville 2006; https://www.fda.gov/OHRMS/DOCKETS/98fr/06d-0344-gdl0001.pdf; last access date October 3 2017
4. L. Richert G. Tuschl C. Abadie N. Blanchard D. Pekthong G. Mantion J. C. Wr andebe S. O. Mueller Use of mRNA expression to detect the induction of drug metabolising enzymes in rat and human hepatocytes Toxicol. Appl. Pharmacol. 235 (2009) 86-96; https://doi.org/10.1016/j.taap.2008.1 J. Skorkovská 1.019 V. Hanušová V. Tománková E. Anzenbacherová B. Lišková P. Anzenbacher
5. P. Matoušková H. Bártíková I. Boušová B. Szotáková J. Martin and L. Skálová Effect of defined green tea extract in various dosage schemes on drug-metabolizing enzymes in mice in vivo J. Funct. Foods 10 (2014) 327-335; https://doi.org/10.1016/j.jff.2014.06.026
6. O. H. Elshenawy and A. O. El-Kadi Modulation of aryl hydrocarbon receptor regulated genes by acute administration of trimethylarsine oxide in the lung kidney and heart of C57BL/6 mice Xenobiotica 45 (2015) 930-943; https://doi.org/10.3109/00498254.2015.1032385
7. O. Hankinson The aryl hydrocarbon receptor complex Annu. Rev. Pharmacol. 35 (1995) 307-340; https://doi.org/10.1146/annurev.pa.35.040195.001515
8. H. M. Korashy and A. O. El-Kadi The role of aryl hydrocarbon receptor in the pathogenesis of cardiovascular diseases Drug Metab. Rev. 38 ( 2006) 4 11-450; https://doi.org/10.1080/03602530600632063
9. C. Abadie-Viollon H. Martin N. Blanchard D. Pekthong P. Bachellier G. Mantion B. Heyd F. Schuler P. Coassolo E. Alexandre and L. Richert Follow-up to the pre-validation of a harmonised protocol for assessment of CYP induction responses in freshly isolated and cryopreserved human hepatocytes with respect to culture format treatment positive reference inducers and incubation conditions Toxicol. In Vitro 24 (2010) 346-356; https://doi.org/10.1016/j.tiv.2009.05.021
10. R. Glockner A. Lieder and A. Lupp Determination of CYP activity in precision-cut liver slices: whether to use intact slices or slice homogenate Anal. Bioanal. Chem. 392 (2008) 1167-1172; https://doi.org/10.1007/s00216-008-2238-y
11. M. Monshouwer G. A. Van’t Klooster S. M. Nijmeijer R. F. Witkamp and A. S. van Miert Characterization of cytochrome P450 isoenzymes in primary cultures of pig hepatocytes Toxicol. In Vitro 12 (1998) 715-723.
12. M. N. Berry G. J. Barritt and A. M. Edwards Isolated Hepatocytes: Preparation Properties and Application Elsevier Amsterdam 1991 Vol. 21.
13. K. Lnenickova E. Prochazkova L. Skalova P. Matouskova H. Bartikova P. Soucek and B. Szotakova Catechins variously affect activities of conjugation enzymes in proliferating and differentiated Caco-2 cells Molecules 21 (2016) e1186 (11 pages); https://doi.org/10.3390/molecules21091186
14. J. J. Cullen M. M. Hinkhouse M. Grady A. W. Gaut J. Liu Y. P. Zhang C. J. Weydert F. E. Domann and L. W. Oberley Dicumarol inhibition of NADPH:quinone oxidoreductase induces growth inhibition of pancreatic cancer via a superoxide-mediated mechanism Cancer Res. 63 (2003) 5513-5520.
15. M. D. Burke and R. T. Mayer Ethoxyresorufin: direct fluorimetric assay of a microsomal O-dealkylation which is preferentially inducible by 3-methylcholanthrene Drug Metab. Dispos. 2 (1974) 583-588.
16. A. Untergasser I. Cutcutache T. Koressaar J. Ye B. C. Faircloth M. Remm and S. G. Rozen Primer3 - new capabilities and interfaces Nucleic Acids Res. 40 (2012) e115 (12 pages); https://doi.org/10.1093/nar/gks596
17. K. J. Livak and T. D. Schmittgen Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method Methods 25 (2001) 402-408; https://doi.org/10.1006/meth.2001.1262
18. Z. Dvorak P. Maurel M. J. Vilarem J. Ulrichova and M. Modriansky Expression and transcriptional activities of nuclear receptors involved in regulation of drug-metabolizing enzymes are not altered by colchiceine: Focus on PXR CAR and GR in primary human hepatocytes Cell Biol. Toxicol. 23 (2007) 63-73; https://doi.org/10.1007/s10565-006-0127-8
19. M. Nishimura A. Koeda E. Suzuki T. Shimizu Y. Kawano M. Nakayama and T. Satoh Effects of prototypical drug-metabolizing enzyme inducers on mRNA expression of housekeeping genes in primary cultures of human and rat hepatocytes Biochem. Bioph. Res. Commun. 346 (2006) 1033-1039; https://doi.org/10.1016/j.bbrc.2006.06.012
20. Y. Korenori S. Tanigawa T. Kumamoto S. Qin Y. Daikoku K. Miyamori M. Nagai and D. X. Hou Modulation of Nrf2/Keap1 system by Wasabi 6-methylthiohexyl isothiocyanate in ARE-mediated NQO1 expression Mol. Nutr. Food Res. 57 (2013) 854-864; https://doi.org/10.1002/mnfr.201200689
21. I. Bousova and L. Skalova Inhibition and induction of glutathione S-transferases by flavonoids: possible pharmacological and toxicological consequences Drug Metab. Rev. 44 (2012) 2 67-286; https://doi.org/10.3109/03602532.2012.713969
22. E. P. Chen L. F. Chen Y. Ji G. Y. Tai Y. H. Wen and H. Ellens A mechanism-based mathematical model of aryl hydrocarbon receptor-mediated CYP1A induction in rats using beta-naphthoflavone as a tool compound Drug Metab. Dispos. 38 (2010) 2278-2285; https://doi.org/10.1124/dmd.110.034421
23. S. Karakurt Modulatory effects of rutin on the expression of cytochrome P450s and antioxidant enzymes in human hepatoma cells Acta Pharm. 66 (2016) 491-502; https://doi.org/10.1515/acph-2016-0046
24. L. G. Higgins and J. D. Hayes Mechanisms of induction of cytosolic and microsomal glutathione transferase (GST) genes by xenobiotics and pro-inflammatory agents Drug Metab. Rev. 43 (2011) 92-137; https://doi.org/10.3109/03602532.2011.567391
25. O. H. Elshenawy and A. O. S. El-Kadi Modulation of aryl hydrocarbon receptor-regulated enzymes by trimethylarsine oxide in C57BL/6 mice: In vivo and in vitro studies Toxicol. Lett. 238 (2015) 17-31; https://doi.org/10.1016/j.toxlet.2015.06.1646
26. I. Bousova J. Hajek J. Drsata and L. Skalova Naturally occurring flavonoids as inhibitors of purified cytosolic glutathione S-transferase Xenobiotica 42 (2012) 872-879; https://doi.org/10.3109/0049-8254.2012.670737
27. D. Brauze K. Fijalkiewicz M. Szaumkessel K. Kiwerska K. Bednarek M. Rydzanicz J. Richter R. Grenman and M. Jarmuz-Szymczak Diversified expression of aryl hydrocarbon receptor dependent genes in human laryngeal squamous cell carcinoma cell lines treated with beta-naphthoflavone Toxicol. Lett. 231 (2014) 99-107; https://doi.org/10.1016/j.toxlet.2014.09.005
28. D. Brauze M. Widerak J. Cwykiel K. Szyfter and W. Baer-Dubowska The effect of aryl hydrocarbon receptor ligands on the expression of AhR AhRR ARNT Hif1 alpha CYP1A1 and NQO1 genes in rat liver Toxicol. Lett. 167 (2006) 212-220; https://doi.org/10.1016/j.toxlet.2006.09.010
29. T. Maier M. Guell and L. Serrano Correlation of mRNA and protein in complex biological samples FEBS Lett. 583 (2009) 3966-3973; https://doi.org/10.1016/j.febslet.2009.10.036