Maternal Perinatal Exposure to Dibutyl Phthalate Promotes Ovarian Dysfunction in Adult Female Offspring via Downregulation of TGF-β2 and TGF-β3

Abstract

Maternal exposure to dibutyl phthalate (DBP) may result in ovarian dysfunction in female offspring. However, the underlying mechanisms remain elusive. Pregnant Sprague-Dawley rats were intraperitoneally injected with different doses of DBP, estradiol, and corn oil from gestational day 7 until the end of lactation. The reproductive characteristics, mRNA, and protein expression of ovaries for the adult female offspring were compared. KGN cells were cultured in vitro with DBP, estrogen receptor antagonist, or ALK-5 inhibitor. Genes, proteins, estradiol, and progesterone expressed by KGN, cell proliferation, and apoptosis were measured respectively. Maternal perinatal exposure to DBP induced prolonged estrous period, increased secondary follicles, significant decreased mRNA, and protein levels of TGF-β2, TGF-β3, and TGF-βRII in ovaries of the adult female offspring, but none difference for serum levels of sex hormones, ovarian TGF-β1, and estrogen receptor. The mRNA levels of LHR, FSHR, and CYP19a in ovaries were also decreased. DBP might decrease the mRNA of TGF-β2, TGF-β3, and TGF-βR II of KGN. DBP can inhibit the mRNA of CYP19 at 24h, which might be blocked by the estrogen receptor antagonist, whose effects were attenuated at 48h. DBP combined with FSH might time-dependently regulate the gene expression of TGF-βR II, inhibitory at 24h, but stimulative at 48h, which could be blocked by the ALK5 inhibitor. However, the protein expressed by KGN was not influenced by DBP. DBP stimulated the proliferation of KGN at 24h, which could be blocked by estrogen receptor antagonist, but attenuated at 48h. The progesterone in culture medium secreted by KGN was decreased by DBP at 24h. Maternal perinatal exposure to DBP induced decreased gene expression of TGF-β signaling and functional proteins in ovaries of the adult female offspring. Molecular cross-talk between estrogen receptor and TGF-β signaling pathway may play role in the mechanism of granulosa dysfunction induced by DBP.