Di-(2-ethylhexyl) phthalate suppresses estradiol and ovulation in cycling rats

Di-(2-ethylhexyl) phthalate (DEHP) is a known reproductive toxicant and a carcinogen in rodent animal models. DEHP may also be a reproductive toxicant in women. Its action in female animal models is undetermined, although its potential to target the ovary has recently been shown. We have identified the ovarian toxicity and target cells of DEHP in the female rat. Adult, regularly cycling Sprague-Dawley rats were dosed daily with 2 g/kg DEHP in corn oil by gavage for 1 to 12 days. Ovarian morphology and serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol, and progesterone levels were analyzed. DEHP exposure resulted in prolonged estrous cycles. Specifically, 35 of 42 DEHP-treated rats had 5- or 6-day cycles and only 7 of 42 had a 4-day cycle compared to 44 of 45 control rats with 4-day cycle lengths. DEHP treatment also suppressed or delayed ovulations by the first proestrus/estrus after metestrus-initiated dosing. Microscopic evaluation of the ovaries determined that 7 of 10 DEHP-exposed rats had not ovulated by vaginal estrus, whereas 13 of 13 control rats had ovulated by vaginal estrus. Thus, DEHP treatment significantly altered natural ovulation times. Preovulatory follicles were also quantitatively smaller in DEHP-exposed rats than in controls because the granulosa cells were smaller. The mean control rat preovulatory follicle granulosa cell area was 16 +/- 3 x 10(3) microns, whereas the mean DEHP-treated rat preovulatory follicle granulosa cell area was 12 +/- 4 x 10(3) microns. DEHP exposure significantly suppressed preovulatory follicle granulosa cell estradiol production over 8 days of exposure. Suppressed serum estradiol levels caused secondary increases in FSH levels and did not stimulate the LH surge necessary for ovulation. Consequently, ovulation did not occur in DEHP-treated rats, although DEHP-treated rats ovulated after treatment with human chorionic hormone. Vaginal lavage cytology from rats treated with DEHP did not detect the ovarian toxicity as shown by histology and hormone analysis. In summary, exposure to DEHP resulted in hypoestrogenic anovulatory cycles and polycystic ovaries in adult female rats.