US EPA

3859077 

Journal Article 

Effects of in vitro exposure to dibutyl phthalate, mono-butyl phthalate, and acetyl tributyl citrate on ovarian antral follicle growth and viability 

Rasmussen, LM; Sen, N; Vera, JC; Liu, X; Craig, ZR 

2017 

Yes 

Biology of Reproduction
ISSN: 0006-3363
EISSN: 1529-7268 

96 

5 

1105-1117 

English 

28486587 

10.1095/biolreprod.116.144691 

WOS:000405440100016 

Dibutyl phthalate (DBP) is present in consumer products and the coating of some oral medications. Acetyl tributyl citrate (ATBC) has been proposed as an alternative to DBP because DBP causes endocrine disruption in animal models. Following ingestion, DBP is converted to its main metabolite mono-butyl phthalate (MBP) which has been detected in >90% of human follicular fluid samples. Previous studies show that DBP reduces the number of antral follicles present in the ovaries of mice. Thus, this study was designed to evaluate the effects of DBP, MBP, and ATBC on in vitro growth and viability of mouse ovarian antral follicles. Antral follicles were isolated from CD-1 females (PND32-37) and treated with vehicle, DBP, MBP, or ATBC (starting at 0.001 and up to 1000 μg/mL for DBP; 24-72 h). Follicle diameter, ATP production, qPCR and TUNEL were used to measure follicle growth, viability, cell cycle and apoptosis gene expression, and cell death-associated DNA fragmentation, respectively. While MBP did not cause toxicity, DBP exposure at ≥10 μg/mL resulted in growth inhibition followed by cytoxicity at ≥500 μg/mL. ATBC increased the number of non-growing follicles at 0.01 μg/mL and did not affect ATP production, but increased TUNEL positive area in treated follicles. Gene expression results suggest that cytotoxicity in DBP-treated follicles occurs via activation of cell cycle arrest prior to follicular death. These findings suggest that concentrations of DBP ≥10 μg/mL are detrimental to antral follicles and that ATBC should be examined further as it may disrupt antral follicle function at low concentrations. 

Adenosine Triphosphate/biosynthesis; Animals; Apoptosis Regulatory Proteins/biosynthesis; Cell Cycle/drug effects; Cell Death/drug effects; Citrates/toxicity; DNA Fragmentation/drug effects; Dibutyl Phthalate/toxicity; Gene Expression/drug effects; Ovarian Follicle/drug effects/growth & development; Plasticizers/toxicity; acetyl tributyl citrate; antral follicle; apoptosis; cell cycle; dibutyl phthalate; endocrine disruptor; mono-butyl phthalate; phthalate substitute; toxicology; 0ZBX0N59RZ; 2286E5R2KE; 8L70Q75FXE