Dual-parent di-(2-ethylhexyl) phthalate (DEHP) exposure: A multigenerational assessment of neurocognitive outcomes in male pups.

Di-(2-ethylhexyl) phthalate (DEHP) is an endocrine disruptor (EDC) of significant public health concern due to its widespread use and potential adverse effects. This study investigated DEHP's multigenerational and dual-parent neurotoxic effects on male rats. The dams of F0 and F1 were exposed to DEHP during the gestation and lactation periods (GD0 to PND21). Meanwhile, F1 fathers were exposed prenatally and postnatally during puberty, maturity, and adulthood to evaluate the impact of paternal DEHP (300 mg/kg) exposure on their offspring. A comprehensive battery of tests was employed, including behavioural assessments (anxiety-like and cognitive behaviour), measurement of oxidative stress, inflammation, and apoptosis biomarkers, analysis of brain neurotransmitter levels, and gene expression and DNA methylation analysis of genes like ER-α, GABA-A5, COX-2, Bcl-2, caspase-3, DNMT1, DNMT3B, and DNMT3A. Results showed that both first- and second-generation pups demonstrate anxiety-like behaviour, working memory impairment, altered brain redox status, the elevation of inflammatory mediators and apoptotic biomarkers, changes in neurotransmitter level, and dysregulation of brain gene expression and DNA methylation, confirmed by brain architecture destruction and elevated apoptotic immunohistochemistry markers. In conclusion, DEHP induced neurotoxicity across multiple generations through the maternal and paternal lineages of male pups.