Hannon Patrick R, Land Katie L, Xu Hong, Akin James W, Curry Thomas E
Department of Obstetrics & Gynecology, University of Kentucky College of Medicine, 800 Rose St, Lexington, KY 40536, USA; Department of Pharmacology and Nutritional Sciences, University of Kentucky College of Medicine, 800 Rose St, Lexington, KY 40536, USA.
Department of Obstetrics & Gynecology, University of Kentucky College of Medicine, 800 Rose St, Lexington, KY 40536, USA.
Reprod Toxicol. 2025 Sep;136:108968. doi: 10.1016/j.reprotox.2025.108968. Epub 2025 May 31.
Exposure to individual phthalates disrupts ovarian function; however, the direct effects of phthalate mixture exposure on ovulation is unknown, especially in women. Human granulosa cells were used to test the hypothesis that exposure to a phthalate mixture (PHTmix; derived from women's urinary phthalate levels) disrupts the function of prostaglandins (PGs), which are vital mediators of ovulation. Additionally, cAMP supplementation was tested as a method to circumvent phthalate toxicity. Granulosa cells from women undergoing in vitro fertilization were acclimated in culture to regain responsiveness to human chorionic gonadotropin (hCG; clinical luteinizing hormone analogue). Granulosa cells were treated with or without hCG, with or without PHTmix (1-500 µg/ml; DMSO=vehicle control), and with or without 8-Br-cAMP (stable cAMP analogue) for 6-36 hr. Exposure to hCG+PHTmix decreased ovulatory PGE and PGF levels when compared to hCG. The mechanism by which the PHTmix decreased PG levels was via decreased synthesis (decreased PTGS2 and PTGES levels) and increased metabolism (increased AKR1C1, AKR1C3, and HPGD levels). Exposure to hCG+PHTmix also impaired PG function by altering levels of PG transporters (ABCC4 and SLCO2A1) and receptors (PTGER2, PTGER3, and PTGFR) when compared to hCG. Supplementation with cAMP in the hCG+PHTmix 500 µg/ml group restored PGE and PGF levels comparable to and beyond hCG control levels. These findings suggest that phthalates inhibit the ovulatory increase in PGs in human granulosa cells via decreased synthesis and increased metabolism. Restored PG levels with cAMP supplementation further establishes a mechanism of toxicity by providing demonstration of a potential cellular target of phthalate-induced ovulatory defects in women.
接触单一邻苯二甲酸盐会扰乱卵巢功能；然而，接触邻苯二甲酸盐混合物对排卵的直接影响尚不清楚，尤其是在女性中。使用人颗粒细胞来检验以下假设：接触邻苯二甲酸盐混合物（PHTmix；源自女性尿液中的邻苯二甲酸盐水平）会扰乱前列腺素（PGs）的功能，而前列腺素是排卵的重要介质。此外，测试了补充环磷酸腺苷（cAMP）作为规避邻苯二甲酸盐毒性的一种方法。对接受体外受精的女性的颗粒细胞进行培养驯化，以恢复其对人绒毛膜促性腺激素（hCG；临床促黄体生成素类似物）的反应性。颗粒细胞分别用或不用hCG、用或不用PHTmix（1 - 500μg/ml；二甲基亚砜=溶剂对照）、用或不用8 - 溴 - cAMP（稳定的cAMP类似物）处理6 - 36小时。与单独使用hCG相比，接触hCG + PHTmix会降低排卵时的前列腺素E（PGE）和前列腺素F（PGF）水平。PHTmix降低PG水平的机制是通过合成减少（PTGS2和PTGES水平降低）和代谢增加（AKR1C1、AKR1C3和HPGD水平增加）。与单独使用hCG相比，接触hCG + PHTmix还会通过改变PG转运体（ABCC4和SLCO2A1）和受体（PTGER2、PTGER3和PTGFR）的水平来损害PG功能。在hCG + PHTmix 500μg/ml组中补充cAMP可使PGE和PGF水平恢复到与hCG对照组相当甚至超过hCG对照组的水平。这些发现表明，邻苯二甲酸盐通过减少合成和增加代谢来抑制人颗粒细胞中PGs的排卵增加。补充cAMP恢复PG水平进一步证实了一种毒性机制，即证明了邻苯二甲酸盐诱导女性排卵缺陷的潜在细胞靶点。
