Department of Biology and Ecology, Faculty of Sciences, University of Novi Sad, 21000 Novi Sad, Serbia.
Clinical Center of Vojvodina, Clinic for Gynecology and Obstetrics, Faculty of Medicine, University of Novi Sad, 21000 Novi Sad, Serbia.
Cells. 2023 Jan 22;12(3):398. doi: 10.3390/cells12030398.
DEHP is an endocrine disruptor that interferes with the function of the female reproductive system. Several studies suggested that DEHP affects steroidogenesis in human and rodent granulosa cells (GC). Some studies have shown that DEHP can also affect the FSH-stimulated steroidogenesis in GC; however, the mechanism by which DEHP affects hormone-challenged steroidogenesis in human GC is not understood. Here, we analyzed the mechanism by which DEHP affects steroidogenesis in the primary culture of human cumulus granulosa cells (hCGC) stimulated with FSH. Cells were exposed to DEHP and FSH for 48 h, and steroidogenesis and the activation of cAMP and ERK1/2 were analyzed. The results show that DEHP decreases FSH-stimulated STAR and CYP19A1 expression, which is accompanied by a decrease in progesterone and estradiol production. DEHP lowers cAMP production and CREB phosphorylation in FSH but not cholera toxin- and forskolin-challenged hCGC. DEHP was not able to decrease steroidogenesis in cholera toxin- and forskolin-stimulated hCGC. Furthermore, DEHP decreases FSH-induced ERK1/2 phosphorylation. The addition of EGF rescued ERK1/2 phosphorylation in FSH- and DEHP-treated hCGC and prevented a decrease in steroidogenesis in the FSH- and DEHP-treated hCGC. These results suggest that DEHP inhibits the cAMP and ERK1/2 signaling pathways, leading to the inhibition of steroidogenesis in the FSH-stimulated hCGC.
DEHP 是一种内分泌干扰物，会干扰女性生殖系统的功能。有几项研究表明，DEHP 会影响人类和啮齿动物颗粒细胞（GC）中的类固醇生成。一些研究表明，DEHP 还可以影响 GC 中 FSH 刺激的类固醇生成；然而，DEHP 影响人 GC 中激素挑战的类固醇生成的机制尚不清楚。在这里，我们分析了 DEHP 影响 FSH 刺激的人卵丘颗粒细胞（hCGC）原代培养中类固醇生成的机制。细胞暴露于 DEHP 和 FSH 48 小时，分析类固醇生成和 cAMP 和 ERK1/2 的激活。结果表明，DEHP 降低了 FSH 刺激的 STAR 和 CYP19A1 表达，这伴随着孕激素和雌二醇产生的减少。DEHP 降低了 FSH 中的 cAMP 产生和 CREB 磷酸化，但不降低霍乱毒素和 forskolin 挑战的 hCGC 中的 cAMP 产生和 CREB 磷酸化。DEHP 不能降低霍乱毒素和 forskolin 刺激的 hCGC 中的类固醇生成。此外，DEHP 降低了 FSH 诱导的 ERK1/2 磷酸化。EGF 的添加挽救了 FSH 和 DEHP 处理的 hCGC 中的 ERK1/2 磷酸化，并防止了 FSH 和 DEHP 处理的 hCGC 中类固醇生成的减少。这些结果表明，DEHP 抑制了 cAMP 和 ERK1/2 信号通路，导致 FSH 刺激的 hCGC 中类固醇生成的抑制。
