槲皮素通过Nrf2/GPX4信号通路缓解玉米赤霉烯酮诱导的猪小肠上皮细胞铁死亡  

Quercetin inhibits zearalenone-induced ferroptosis of porcine small intestinal epithelial cells via regulating Nrf2/GPX4 signaling pathway

作　　者：陈海燕[1] 卢慧芳[1] 曹智高[1] 孙攀峰[1] 刘守铉 宋超 CHEN Haiyan;LU Huifang;CAO Zhigao;SUN Panfeng;LIU Shouxuan;SONG Chao(College of Modern Animal Husbandry,Henan Vocational College of Agriculture,Zhengzhou 451450,China;College of Veterinary Medicine,Henan University of Animal Husbandry and Economy,Zhengzhou 457499,China)

机构地区：[1]河南农业职业学院现代牧业学院,河南郑州451450 [2]河南牧业经济学院动物医药学院,河南郑州457499

出　　处：《中国兽医学报》2025年第3期527-534,593,共9页Chinese Journal of Veterinary Science

基　　金：河南省科技攻关计划资助项目(242102110012);河南省自然科学基金资助项目(232300420218)。

摘　　要：基于核因子E2相关因子2(Nrf2)/谷胱甘肽过氧化物酶4(GPX4)信号通路研究槲皮素抑制玉米赤霉烯酮(zearalenone,ZEN)诱导IPEC-J2猪小肠上皮细胞铁死亡的作用机制。体外培养IPEC-J2细胞,ZEN处理(25mg/L)的同时给予不同浓度槲皮素干预(10、20、40μmol/L)。生化法检测各组细胞培养液上清中乳酸脱氢酶(LDH)、总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)活性、谷胱甘肽(GSH)和丙二醛(MDA)水平;荧光探针检测细胞中Fe^(2+)、活性氧(ROS)和脂质过氧化水平;Western blot检测Nrf2、长链酯酰辅酶A合成酶4(ACSL4)和GPX4蛋白表达水平。将IPEC-J2细胞分为对照组、ZEN组、槲皮素组和ML385(Nrf2抑制剂)+槲皮素组,除对照组外其余各组均接受ZEN处理,并采用槲皮素和ML385进行干预,检测Nrf2/GPX4通路相关蛋白表达和铁死亡指标(LDH、Fe^(2+)、MDA和GSH)变化。结果显示,与对照组比较,ZEN组细胞存活率、T-AOC及GSH、SOD水平、Nrf2和GPX4蛋白表达降低(P<0.05),细胞LDH释放率、Fe^(2+)及ROS、脂质过氧化、MDA水平及ACSL4蛋白表达升高(P<0.05)。与ZEN组比较,槲皮素组细胞存活率、细胞T-AOC、SOD及GSH水平、Nrf2和GPX4蛋白表达升高(P<0.05),细胞LDH释放率、Fe^(2+)及ROS、脂质过氧化、MDA水平及ACSL4蛋白表达降低(P<0.05)。与槲皮素组比较,ML385+槲皮素组细胞Nrf2及GPX4蛋白表达和GSH水平降低(P<0.05),细胞LDH释放率、Fe^(2+)及MDA水平升高(P<0.05)。综上,槲皮素能够抑制ZEN诱导的IPEC-J2细胞铁死亡,其作用机制可能与其激活Nrf2/GPX4信号通路有关。The aim of this study was to investigate the mechanism of quercetin in inhibiting the ferroptosis 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(10,20,40μmol/L).Biochemical methods were used to detect the levels of lactate dehydrogenase(LDH)in the cell culture medium supernatant,total antioxidant capacity(T-AOC),superoxide dismutase(SOD),glutathione(GSH),and malondialdehyde(MDA).Fluorescence probes were used to detect the levels of Fe^(2+),reactive oxygen species(ROS),and lipid peroxidation.Western blot was performed to detect 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(Nrf2inhibitor)+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 pathway-related proteins and ferroptosis-related indexes(LDH,Fe^(2+),MDA,and GSH)were detected.Compared with the control group,IPEC-J2 cells in the ZEN group exhibited a decrease in cell viability,T-AOC,and GSH,SOD levels,Nrf2,and GPX4 protein expressions(P<0.05),while LDH release rate,Fe^(2+)and ROS,lipid peroxidation,MDA levels,and ACSL4 protein expression decreased in the ZEN group(P<0.05).Compared with ZEN group,the cell viability,the levels of TAOC,SOD and GSH,the protein expression of Nrf2 and GPX4 in quercetin groups were 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.

关 键 词：玉米赤霉烯酮 槲皮素 猪小肠上皮细胞 铁死亡 Nrf2/GPX4 

分 类 号：S859.7[农业科学—临床兽医学] S859.8[农业科学—兽医学]