Table4_Di-(2-ethylhexyl) phthalate exposure induces liver injury by promoting ferroptosis via downregulation of GPX4 in pregnant mice.XLSX

As one kind of endocrine disrupting chemical, di-(2-ethylhexyl) phthalate (DEHP) has been reported to cause liver dysfunction in epidemiological and experimental studies. Abnormal liver function in pregnancy is associated with adverse maternal and perinatal outcomes. Few studies have investigated the potential effect of gestational DEHP exposure on the liver in pregnant mice, and the underlying mechanisms remain unclear. In the present study, pregnant ICR mice were exposed to doses (0, 500, 1,000 mg/kg/day) of DEHP in the presence or absence of 5 mg/kg/day ferrostatin-1 (Fer-1, ferroptosis inhibitor) by oral gavage from gestation day 4 to day 18. HepG2 cells were exposed to different doses of monoethylhexyl phthalate (MEHP, a major metabolite of DEHP) in vitro. Hepatic function and pathologic changes were observed. Oxidative stress, iron metabolism, and ferroptosis-related indicators and genes were evaluated both in vivo and in vitro. The results showed that gestational DEHP exposure induced disordered liver function and hepatocyte morphology changes in pregnant mice, along with increased malondialdehyde (MDA) and Fe2+ content and decreased glutathione (GSH) levels. The expression levels of the selected ferroptosis-related genes Slc7a11, Gpx4, and Nfr2 were significantly decreased, and Ptgs2 and Lpcat3 were significantly increased. Notably, Fer-1 attenuated DEHP-induced liver injury and ferroptosis. Furthermore, MEHP exhibited a synergistic effect with RSL3 (a GPX4 inhibitor) in promoting ferroptosis in vitro. Taken together, the results demonstrated that DEHP induced liver injury and ferroptosis in pregnant mice, probably by inhibiting the GPX4 pathway through lipid peroxidation and iron accumulation.

作为一种内分泌干扰化学物质，对二(2-乙基己基)邻苯二甲酸（DEHP）的报道指出，其在流行病学和实验研究中可导致肝功能异常。妊娠期间的肝脏功能异常与母体和围产期不良结局相关。关于妊娠期DEHP暴露对孕鼠肝脏潜在影响的调查为数不多，其作用机制尚不明确。在本研究中，从妊娠第4天至第18天，对ICR孕鼠进行口服灌胃，暴露于DEHP的剂量分别为（0、500、1,000 mg/kg/天），同时或在5 mg/kg/天铁死亡抑制剂-1（Fer-1）的辅助下进行。体外培养的HepG2细胞则暴露于不同剂量的单乙基己基邻苯二甲酸（MEHP，DEHP的主要代谢物）。观察了肝功能及病理变化。在体内和体外评估了氧化应激、铁代谢和铁死亡相关指标及基因。结果显示，妊娠期DEHP暴露导致孕鼠肝脏功能紊乱和肝细胞形态改变，同时增加了丙二醛（MDA）和Fe2+含量，降低了谷胱甘肽（GSH）水平。选定的铁死亡相关基因Slc7a11、Gpx4和Nfr2的表达水平显著降低，而Ptgs2和Lpcat3的表达水平显著升高。值得注意的是，Fer-1减轻了DEHP诱导的肝脏损伤和铁死亡。此外，MEHP在体外与RSL3（一种GPX4抑制剂）表现出协同促进铁死亡的效果。综上所述，研究结果表明，DEHP通过抑制GPX4通路，通过脂质过氧化和铁积累，可能诱导孕鼠肝脏损伤和铁死亡。