Comparison of cytochrome P450 activity and mRNA expression in canine vs. human hepatocytes after acetaminophen, diclofenac, or valproic acid exposure.

IMPORTANCE

Although extensive studies have been conducted on cytochrome P450 (CYP) enzymes in rodents and humans, research on canine CYP enzymes is limited. The lack of species-specific metabolic research on dogs presents a major challenge in predicting toxicity and adverse drug reactions.

OBJECTIVE

This study aimed to examine the interspecies differences in CYP enzyme activity and mRNA expression between canine and human hepatocytes following treatment with acetaminophen (AAP), diclofenac (Dic), or valproic acid (VPA).

METHODS

We determined the 24-h exposure half-maximal inhibitory concentration (IC₅₀) values of AAP, Dic, and VPA in canine and human hepatocytes. Based on these IC₅₀ concentrations, we compared drug-induced alterations in various parameters, including immunocytochemistry, transcriptomic profiles (RNA-seq), and CYP activity, to assess changes at the gene and protein levels.

RESULTS

AAP and VPA increased CYP2J2 mRNA expression by 4.7- and 7.76-fold, respectively, whereas Dic increased CYP1A1 mRNA expression by 14.25-fold in canine hepatocytes. AAP, VPA, and Dic decreased CYP26B1 mRNA expression in canine hepatocytes by 0.03-, 0.12-, and 0.17-fold, respectively. Dic and VPA increased CYP1A1 mRNA expression by 5.53- and 6.66-fold, respectively, whereas AAP, VPA, and Dic decreased CYP4F22 mRNA expression by 0.03-, 0.13-, and 0.13-fold, respectively, in human hepatocytes.

CONCLUSIONS AND RELEVANCE: The observed differences between species in CYP activity and mRNA levels in response to drug exposure highlight the importance of accurate and precise experimental models for the development of new medications.