Plastic additives affect estrogenic pathways and lipid metabolism in precision - cut - liver slices in Atlantic cod (Gadus morhua)

The overall aim of the present study was to determine if exposure to three high volume plastic additives, including diethylhexyl phthalate (DEHP), bisphenol A (BPA) and benzotriazoles (BT), have the potential to promote adverse effects in Atlantic cod (G. morhua). Ex vivo precision cut - liver slices (PCLS) from six male juvenile Atlantic cod were exposed to four concentrations of mono-(2-ethylhexyl)-phthalate (MEHP, the main metabolite of DEHP), BPA and BT both singly and in mixtures ranging from 0.1 to 100 μM (MEHP), 0.022-22 μM (BPA) and 0.042-42 μM (BT). Histology and transmission electron microscopy (TEM) were used to assess pathological changes and ultrastructure of the exposed liver tissue. Vitellogenin (Vtg) produced by the hepatic tissue was analyzed using ELISA, and the transcription levels of selected biomarker genes (vtg1, esr1, cyp1a, scdb, aclya, fabp1a, acox1, hnf4a and cebp) were measured using Quantitative real-time polymerase chain reaction (Q-PCR). An estrogenic effect was observed with a significant upregulation of the vtg1 and esr1 genes and increase in Vtg protein synthesis following exposure to BPA and a mixture of the selected compounds. The hnf4a showed a significant downregulation following mixture exposure, where the BPA was suspected to be the main driver for this response although not inducing a significant downregulation in the single component exposure. There was no significant difference between the mixture exposure and the individual compound exposures, nevertheless a tendency of an antagonistic mixture effect for the biomarkers of estrogenic effect (vtg1, esr1 and Vtg), and possibly synergistic or additive effect on the lipid metabolism related gene hnf4a, warrants further investigation.