PEDV infection induces ferroptosis in Vero cells via an ACSL4-mediated lipid peroxidation pathway

Abstract Porcine epidemic diarrhea virus (PEDV) is a highly contagious viral pathogen causing severe economic losses in the swine industry. However, the underlying mechanisms by which PEDV induces host cell death are largely unknown, leading to an initial stage of drug development against PEDV. In this study, we investigated the role of ferroptosis, a non-apoptotic form of programmed cell death, in PEDV-infected cells. The experiments were divided into four groups: a control group, a PEDV group, an Erastin positive control group, and a Liprostatin negative control group. Levels of GSH, ROS, Fe3+, and cell viability were evaluated using ELISA test kits. Fluorescence microscopy was employed to assess Fe2+ aggregation, while flow cytometry was utilized to measure lipid peroxide levels. The mRNA transcript levels of key genes involved in the ferroptosis pathway -ACSL4, GPX4, ALOX15, and LPCAT3 - were determined by quantitative reverse transcription PCR. Compared to the control group, the PEDV group exhibited a significant decrease in GSH levels and a gradual reduction in Fe3+ levels. Furthermore, the PEDV group showed a substantial increase in ROS release and a corresponding decrease in cell viability relative to the control group. The expression levels of ACSL4, ALOX15 and LPCAT3 mRNA were significantly elevated in the PEDV group. Additionally, the protein expression levels of ACSL4, ALOX15 and LPCAT3 also increased progressively. In conclusion, our findings suggest that PEDV can induce iron prolapse in Vero cells via the ACSL4-mediated lipid peroxidation pathway, which furthers our knowledge of PEDV and provides a theoretical basis for drug development.