HFPO-TA induces liver damage and ferroptosis in zebrafish by disrupting iron metabolism homeostasis.

Hexafluoropropylene oxide trimeric acid (HFPO-TA) is a novel alternative to perfluorooctanoic acid (PFOA) with similar toxicity to PFOA. However, whether HFPO-TA causes ferroptosis is currently unknown. In this experiment, we systematically investigated the induction of zebrafish embryonic liver ferroptosis by HFPO-TA to cause liver injury. The liver morphology and the extent of inflammatory damage were then visualized by Sudan Black B and H&E staining, as well as through observations of Tg (fabp10a: DsRed), Tg (lyz: DsRed) and Tg (Xla.Eef1a1: mlsEGFP) transgenic zebrafish. Differentially expressed genes associated with ferroptosis were identified using transcriptomics data previously done in the lab, and from these, three were selected as receptors for molecular docking with HFPO-TA, and both receptors and ligands were able to dock successfully. We then observed zebrafish under the microscope using ROS, DPPP and FerroOrange staining, respectively. The expression of key genes was verified by RT-qPCR. The results showed that HFPO-TA could lead to a significant increase in the levels of ROS, lipid peroxides and divalent iron ions, liver inflammation and injury, and a trend of elevated differential genes such as tfa and tfr1a. We then validated the ferroptosis pathway using an agonist (RSL3) and an inhibitor (Fer-1), and found that the inhibitor was able to ameliorate HFPO-TA-induced inflammatory and ferroptosis responses, whereas the agonist did the opposite. In conclusion, the ability of HFPO-TA to induce ferroptosis, which leads to liver injury provides new insights into the mechanism of HFPO-TA toxicity.