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

Fan Zhang; Hualong Zhen; Hengshun Cheng; Fengying Hu; Yunfei Jia; Binbin Huang; Minmin Jiang

doi:10.3389/fcell.2022.1014243

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

Front Cell Dev Biol. 2022 Nov 10:10:1014243. doi: 10.3389/fcell.2022.1014243. eCollection 2022.

Authors

Fan Zhang¹, Hualong Zhen¹, Hengshun Cheng¹, Fengying Hu¹, Yunfei Jia¹, Binbin Huang^{2

3

4}, Minmin Jiang¹

Affiliations

¹ Department of Occupational Health and Environmental Health, School of Public Health, Anhui Medical University, Hefei, China.
² MOE Key Laboratory of Population Health Across Life Cycle, Department of Maternal, Child and Adolescent Health, School of Public Health, Anhui Medical University, Hefei, China.
³ NHC Key Laboratory of Study on Abnormal Gametes and Reproductive Tract, Department of Maternal, Child and Adolescent Health, School of Public Health, Anhui Medical University, Hefei, China.
⁴ Anhui Provincial Key Laboratory of Population Health and Aristogenics, Department of Maternal, Child and Adolescent Health, School of Public Health, Anhui Medical University, Hefei, China.

Abstract

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.

Keywords: DEHP; endocrine disrupting chemical; ferroptosis; liver injury; pregnancy.