The fungicide difenoconazole alters mRNA expression levels of human CYP3A4 in HepG2 cells

The fungicide difenoconazole has been frequently detected in agricultural products, soils and surface waters, causing increasingly public concern due to toxicological properties. Although systemic triazole fungicides can inhibit the enzymatic activity of many CYP450 isozymes, how difenoconazole affects the human CYP3A4 still remains largely unknown. We thus investigated the effect of difenoconazole on normal mRNA expression and protein expression of human CYP3A4 by real-time quantitative PCR and western blot, respectively. Results show that the exposure to difenoconazole from 0.01 to 0.5 μM for 24 h down-regulates mRNA expression levels of human CYP3A4 in HepG2 cells. We also found that difenoconazole could inhibit the enzymatic activity of human CYP3A4 in a concentration-dependent mode. The IC₅₀ of difenoconazole for inhibition of CYP3A4 activity is 0.41 µM, showing a stronger inhibitor in comparison with ketoconazole. Overall, our findings indicate the potential risk of difenoconazole for the disruption of human CYP3A4.