Transgenerational Effects of Di(2-Ethylhexyl) Phthalate on Anogenital Distance, Sperm Functions and DNA Methylation in Rat Offspring.

Li-Ping Huang,Kuo-Hsin Lee,Jia-Ying Jhong,Yue-Leon Guo,Hsin-Pao Chen,Ping-Chi Hsu

doi:10.3390/ijms22084131

Li-Ping Huang, Kuo-Hsin Lee + Show 4 more

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https://doi.org/10.3390/ijms22084131

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Abstract

Di(2-ethylhexyl) phthalate (DEHP) is widely used as a plasticizer in the manufacture of polyvinylchloride plastics and has been associated with concerns regarding male reproductive toxicity. In this study, we hypothesized that maternal exposure to DEHP induces transgenerational inheritance of adult-onset adverse reproductive outcomes through the male germline in the F1, F2, and F3 generations of male offspring. Pregnant rats were treated with 5 or 500 mg of DEHP/kg/day through gavage from gestation day 0 to birth. The offspring body weight, anogenital distance (AGD), anogenital index (AGI), sperm count, motility, and DNA fragmentation index (DFI) were measured for all generations. Methyl-CpG binding domain sequencing was performed to analyze sperm DNA methylation status in the F3. DEHP exposure at 500 mg/kg affected AGD, AGI, sperm count, mean DFI, and %DFI in the F1; AGD, sperm count, and mean DFI in the F2; and AGD, AGI, mean DFI, and %DFI in the F3. DEHP exposure at 5 mg/kg affected AGD, AGI, sperm count, and %DFI in the F1; sperm count in the F2; and AGD and AGI in F3. Compared with the control group, 15 and 45 differentially hypermethylated genes were identified in the groups administered 5 mg/kg and 500 mg/kg DEHP, respectively. Moreover, 130 and 6 differentially hypomethylated genes were observed in the groups administered 5 mg/kg and 500 mg/kg DEHP. Overall, these results demonstrated that prenatal exposure to DEHP caused transgenerational epigenetic effects, which may explain the observed phenotypic changes in the male reproductive system.

Highlights

Di(2-ethylhexyl) phthalate (DEHP), a well-known endocrine-disrupting chemical (EDC), is widely used as a plasticizer in the manufacture of polyvinylchloride (PVC) plastics and has been associated with concerns about antiandrogenic activity and male reproductive toxicity [1,2,3]
These findings indicate that DEHP exposure may disrupt the development of human male genitals; studies examining the concentrations of phthalate metabolites in urine or amniotic fluid and Anogenital distance (AGD) in male offspring have yielded inconsistent results
In the F1 generation, prenatal exposure to the vehicle, 5 mg/kg/day DEHP, or 500 mg/kg/day DEHP from gestational day (GD) 0 to GD 18 did not result in significant differences in the body weights of offspring rats (Figure 1A)

Summary

Introduction

Di(2-ethylhexyl) phthalate (DEHP), a well-known endocrine-disrupting chemical (EDC), is widely used as a plasticizer in the manufacture of polyvinylchloride (PVC) plastics and has been associated with concerns about antiandrogenic activity and male reproductive toxicity [1,2,3]. DEHP and its metabolites are often detected in human urine and amniotic fluid, which underscores its risk as a toxic hazard [4,5] and indicates potential effects on fetal development [6,7,8]. No significant association was observed between the phthalate level in the amniotic fluid or urine of Taiwanese women and AGD in their 33 male newborns [14]. These findings indicate that DEHP exposure may disrupt the development of human male genitals; studies examining the concentrations of phthalate metabolites in urine or amniotic fluid and AGD in male offspring have yielded inconsistent results

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