Induction of cytochrome P450 enzymes in rat liver by two conazoles, myclobutanil and triadimefon

Abstract

This study was undertaken to examine the inductive effects of two triazole antifungal agents, myclobutanil and triadimefon, on the expression of hepatic cytochrome P450 (CYP) genes and on the activities of CYP enzymes in male Sprague–Dawley rats. Rats were dosed with the conazoles at three dose levels by gavage for 14 days: myclobutanil (150, 75, and 10 mg kg⁻¹ body weight day⁻¹); triadimefon (115, 50, and 10 mg kg⁻¹ body weight day⁻¹), which included their maximum tolerated dose levels (MTD). Both myclobutanil and triadimefon significantly induced pentoxyresorufin O-depentylase activities at their MTD levels: myclobutanil, 8.1-fold at 150 mg kg⁻¹ body weight day⁻¹; and triadimefon, 18.5-fold at 115 mg kg⁻¹ body weight day⁻¹. Benzyloxyresorufin O-debenzylase activities were similarly increased: myclobutanil, 13.3-fold; triadimefon, 27.7-fold. Quantitative real-time reverse-transcription polymerase chain reaction assays were used to characterize the mRNA expression of specific CYP genes induced by these two conazoles. Myclobutanil and triadimefon treatment at their MTD levels significantly increased rat hepatic mRNA expression of CYP2B1 (14.3- and 54.6-fold), CYP3A23/3A1 (2.2- and 7.3-fold), and CYP3A2 (1.5- and 1.7-fold). Western immunoblots of rat hepatic microsomal proteins identified significantly increased levels of CYP isoforms after myclobutanil or triadimefon treatment at their MTD levels: CYP2B1/2 (4.8- and 5.3-fold), and CYP3A1 (2.2- and 2.9-fold). Triadimefon also increased CYP3A2 immunoreactive protein levels 1.8-fold. These results indicate that triadimefon and myclobutanil, like other triazole-containing conazoles, induced CYP2B and CYP3A families of cytochromes in rat liver.

Key Words::

• Myclobutanil
• triadimefon
• polymerase chain reaction (PCR)
• cytochrome P450 (CYP)
• alkoxyresorufin O-dealkylation
• Western blots
• 7-pentoxyresorufin O-depentylase (PROD)
• 7-benzyloxyresorufin O-debenzylase (BROD)
• 7-methoxyresorufin O-demethylase (MROD)
• 7-ethoxyresorufin O-deethylase (EROD)
• CYP2B1
• CYP3A23/3A1
• CYP3A2

Acknowledgements

The research described herein was reviewed by the National Health and Environmental Effects Research Laboratory, US Environmental Protection Agency, and approved for publication. Approval does not signify that the contents necessarily reflect the views and policies of the Agency, nor does mention of trade names or commercial products constitute endorsement or recommendation for use.