Insights into the effects of difenoconazole on the livers in male mice at the biochemical and transcriptomic levels.

Difenoconazole (DFZ) is a broad-spectrum triazole fungicide, that is extensively used in agriculture. Studies have shown that residues of DFZ and other fungicides have toxic effects on nontarget organisms. However, its hepatoxicity in mammals remains unclear. Here, we characterized the toxic hepatic effects in male C57BL/6 mice exposed to 30 and 100 mg/kg bw DFZ for 14 and 56 days, respectively. The results revealed that DFZ could increase the relative liver weights, however, the relative fat and spleen weights decreased. More importantly, DFZ exposure changed the hepatic morphology and induced hepatic oxidative stress. Gene expression analysis suggested that DFZ could induce a glycolipid metabolism disorder. Moreover, hepatic transcriptomic analysis revealed the effects of DFZ exposure on the transcriptional levels of various genes, and enrichment analysis of differentially expressed genes (DEGs) showed that energy metabolism and immune-associated pathways were mainly affected. We validated the results from transcriptomic analysis and found that some key genes related to energy metabolism were affected. In addition, flow cytometry showed that the CD3+/CD4+ and CD3+ /CD8+ levels declined in the spleen of mice. Taken together, these findings combined with transcriptome analysis highlighted that DFZ caused different endpoints in the liver, which could provide more evidence for investigating the toxic effects of DFZ in mammals.