Gestational and lactational exposure to DEHP triggers ACSL4/TFR-mediated hippocampal neuronal ferroptosis via YAP activation: Implication for the neurocognitive disorders in male offspring

Abstract

Di-(2-ethylhexyl) phthalate (DEHP) is one of the most extensively used phthalate and poses a public health concern. Perinatal exposure to DEHP has been shown to cause neurodevelopmental abnormalities and neurobehavioral disorders in offspring. However, the precise molecular mechanism has not yet been fully elucidated. In this study, pregnant C57BL/6 mice were exposed to DEHP from gestation to weaning. By RNA sequencing and animal experiments, ferroptosis has been identified as the key pathologic process contributing to DEHP-induced hippocampal injury in adult male offspring. In vitro results also showed that Ferrostatin-1 (Fer-1) effectively ameliorated Mono-(2-ethylhexyl) phthalate (MEHP) -induced cell survival via the inhibiting ferroptosis in HT22 cells. Consistently, we found that the expression of ACSL4 and TFR was significantly up-regulated in offspring hippocampi and MEHP-exposed HT22 neurons. However, silencing ACSL4 or knockdown TFR relieved MEHP-induced generation of lipid ROS and cellular iron accumulation, thereby blocking ferroptosis. Mechanistically, ACSL4/TFR-mediated ferroptosis seemed to be a Yes-associated protein (YAP) dependent via TEA domain transcription factor 4 in HT22 neurons. Importantly, treatment with Fer-1, rosiglitazone, and Deferoxamine effectively rescued DEHP-evoked cognitive decline in adult male offspring. Our findings certified that gestational and lactational exposure to DEHP provoked ACSL4/TFR-mediated hippocampal neuronal ferroptosis via YAP activation.