The aim of this study was to investigate the mechanism of quercetin in inhibiting the fer-roptosis induced by zearalenone(ZEN)via mediating the nuclear factor erythroid-2-related factor 2(Nrf2)/glutathione peroxidase 4(GPX4)signaling pathway.IPEC-J2 cells were cultured in vitro and treated with ZEN(25 mg/L)and different concentrations of quercetin(10,20,40 μmol/L).Bi-ochemical methods were used to detect the levels of lactate dehydrogenase(LDH)in the cell cul-ture medium supernatant,total antioxidant capacity(T-AOC),superoxide dismutase(SOD),glu-tathione(GSH),and malondialdehyde(MDA).Fluorescence probes were used to detect the levels of Fe2+,reactive oxygen species(ROS),and lipid peroxidation.Western blot was performed to de-tect the expression levels of Nrf2,long chain acyl CoA synthetase 4(ACSL4),and GPX4.The IPEC-J2 cells were divided into the control group,ZEN group,quercetin group,and ML385(Nrf2 inhibitor)+quercetin group for further analysis.Except for the control group,the other groups were treated with ZEN in the prescence of quercetin and ML385.The changes of Nrf2/GPX4 path-way-related proteins and ferroptosis-related indexes(LDH,Fe2+,MDA,and GSH)were detected.Compared with the control group,IPEC-J2 cells in the ZEN group exhibited a decrease in cell via-bility,T-AOC,and GSH,SOD levels,Nrf2,and GPX4 protein expressions(P＜0.05),while LDH release rate,Fe2+and ROS,lipid peroxidation,MDA levels,and ACSL4 protein expression de-creased in the ZEN group(P＜0.05).Compared with ZEN group,the cell viability,the levels of T-AOC,SOD and GSH,the protein expression of Nrf2 and GPX4 in quercetin groups were increased(P＜0.05),while the LDH release rate,the levels of Fe2+,ROS,lipid peroxidation,and MDA as well as the protein expression of ACSL4 were decreased(P＜0.05).Compared with the quercetin group,the protein expression of Nrf2 and GPX4 and the level of GSH in ML385+quercetin group reduced(P＜0.05),the LDH release rate and the levels of Fe2+and MDA increased(P＜0.05).In summary,quercetin could inhibit ZEN-induced ferroptosis of IPEC-J2 cells,and its mechanism may be related to the induction of Nrf2/GPX4 signaling pathway.

The aim of this study was to investigate the mechanism of quercetin in inhibiting the fer-roptosis induced by zearalenone(ZEN)via mediating the nuclear factor erythroid-2-related factor 2(Nrf2)/glutathione peroxidase 4(GPX4)signaling pathway.IPEC-J2 cells were cultured in vitro and treated with ZEN(25 mg/L)and different concentrations of quercetin(10,20,40 μmol/L).Bi-ochemical methods were used to detect the levels of lactate dehydrogenase(LDH)in the cell cul-ture medium supernatant,total antioxidant capacity(T-AOC),superoxide dismutase(SOD),glu-tathione(GSH),and malondialdehyde(MDA).Fluorescence probes were used to detect the levels of Fe2+,reactive oxygen species(ROS),and lipid peroxidation.Western blot was performed to de-tect the expression levels of Nrf2,long chain acyl CoA synthetase 4(ACSL4),and GPX4.The IPEC-J2 cells were divided into the control group,ZEN group,quercetin group,and ML385(Nrf2 inhibitor)+quercetin group for further analysis.Except for the control group,the other groups were treated with ZEN in the prescence of quercetin and ML385.The changes of Nrf2/GPX4 path-way-related proteins and ferroptosis-related indexes(LDH,Fe2+,MDA,and GSH)were detected.Compared with the control group,IPEC-J2 cells in the ZEN group exhibited a decrease in cell via-bility,T-AOC,and GSH,SOD levels,Nrf2,and GPX4 protein expressions(P＜0.05),while LDH release rate,Fe2+and ROS,lipid peroxidation,MDA levels,and ACSL4 protein expression de-creased in the ZEN group(P＜0.05).Compared with ZEN group,the cell viability,the levels of T-AOC,SOD and GSH,the protein expression of Nrf2 and GPX4 in quercetin groups were increased(P＜0.05),while the LDH release rate,the levels of Fe2+,ROS,lipid peroxidation,and MDA as well as the protein expression of ACSL4 were decreased(P＜0.05).Compared with the quercetin group,the protein expression of Nrf2 and GPX4 and the level of GSH in ML385+quercetin group reduced(P＜0.05),the LDH release rate and the levels of Fe2+and MDA increased(P＜0.05).In summary,quercetin could inhibit ZEN-induced ferroptosis of IPEC-J2 cells,and its mechanism may be related to the induction of Nrf2/GPX4 signaling pathway.