Prenatal exposure to DEHP and high-fat diet synergistically disrupts gonadal function in male mice

Abstract Prenatal exposure to Di(2-ethylhexyl) phthalate (DEHP) impairs the reproductive system and causes fertility defects in male offspring. Additionally, high-fat diet (HF) is a risk factor for reproductive disorders in males. In this study, we tested the hypothesis that prenatal exposure to a physiologically relevant dose of DEHP in conjunction with HF diet synergistically impacts reproductive function and fertility in male offspring. Female mice were fed a control or HF diet 7 days prior to mating and until their litters were weaned on postnatal day 21. Pregnant dams were exposed to DEHP (20 μg/kg/day) or vehicle from gestational day 10.5 until birth. The male offspring’s gross phenotype, sperm quality, serum hormonal levels, testicular histopathology, and testicular gene expression pattern were analyzed. Male mice born to dams exposed to DEHP + HF had smaller testes, epididymides, and shorter anogenital distance compared to those exposed to HF or DEHP alone. Sperm analysis revealed that DEHP+HF mice had lower sperm concentration and motility compared to DEHP mice. Moreover, DEHP+HF mice had more apoptotic germ cells, fewer Leydig cells, and lower serum testosterone levels than DEHP mice. Further, testicular mRNA expression of Dnmt1 and Dnmt3a was two to eight-fold higher than in DEHP mice by qPCR, suggesting that maternal HF diet and prenatal DEHP exposure additively impact gonadal function by altering the degree of DNA methylation in the testis. These results suggest that the combined exposure to DEHP and HF synergistically impairs reproductive function in male offspring, greater than exposure to DEHP or HF diet alone.