Metabolic signatures of bisphenol A and genistein in Atlantic salmon liver cells.

Screening has revealed that aquafeeds with high inclusion of plant material may contain small amounts of endocrine disrupting agricultural pesticides. In this work, bisphenol A (BPA) and genistein (GEN) were selected as model endocrine disrupting toxicants with impact on DNA methylation in fish. Atlantic salmon hepatocytes were exposed in vitro to four concentrations of BPA and GEN (0.1, 1.0, 10 and 100 μM) for 48 h. Toxicity endpoints included cytotoxicity, global DNA methylation, targeted transcriptomics and metabolomic screening (100 μM). GEN was not cytotoxic in concentrations up to 100 μM, whereas one out of two cell viability assays indicated a cytotoxic response to 100 μM BPA. Compared to the control, significant global DNA hypomethylation was observed at 1.0 μM BPA. Both compounds upregulated cyp1a1 transcription at 100 μM, while estrogenic markers esr1 and vtg1 responded strongest at 10 μM. Dnmt3aa transcription was downregulated by both compounds at 100 μM. Metabolomic screening showed that BPA and GEN resulted in significant changes in numerous biochemical pathways consistent with alterations in carbohydrate metabolism, indicating perturbation in glucose homeostasis and energy generation, and glutamate metabolism. Pathway analysis showed that while the superpathway of methionine degradation was among the most strongly affected pathways by BPA, GEN induced changes to uridine and pyrimidine biosynthesis. In conclusion, this mechanistic study proposes metabolites associated with glucose and glutamate metabolism, glucuronidation detoxification, as well as cyp1a1, vtg1, esr1, ar, dnmt3aa, cdkn1b and insig1 as transcriptional markers for BPA and GEN exposure in fish liver cells.