University of Novi Sad, Faculty of Sciences, Department of Biology and Ecology, Serbia.
University of Novi Sad, Faculty of Sciences, Department of Biology and Ecology, Serbia.
Reprod Toxicol. 2023 Sep;120:108452. doi: 10.1016/j.reprotox.2023.108452. Epub 2023 Aug 2.
Di(2-ethylhexyl) phthalate (DEHP) is an endocrine disruptor that exerts anti-steroidogenic effects in human granulosa cells; however, the extent of this effect depends on the concentration of DEHP and granulosa cell models used for exposure. The objective of this study was to identify the effects of low- and high-dose DEHP exposure in human granulosa cells. We exposed human granulosa cell line HGrC1 to 3 nM and 25 μM DEHP for 48 h. The whole genome transcriptome was analyzed using the DNBSEQ sequencing platform and bioinformatics tools. The results revealed that 3 nM DEHP did not affect global gene expression, whereas 25 µM DEHP affected the expression of only nine genes in HGrC1 cells: ABCA1, SREBF1, MYLIP, TUBB3, CENPT, NUPR1, ASS1, PCK2, and CTSD. We confirmed the downregulation of ABCA1 mRNA and SREBP-1 protein (encoded by the SREBF1 gene), both involved in cholesterol homeostasis. Despite these changes, progesterone production remained unaffected in low- and high-dose DEHP-exposed HGrC1 cells. The high concentration of DEHP decreased the levels of ABC1A mRNA and SREBP-1 protein and prevented the upregulation of STAR, a protein involved in progesterone synthesis, in forskolin-stimulated HGrC1 cells; however, the observed changes were not sufficient to alter progesterone production in forskolin-stimulated HGrC1 cells. Overall, this study suggests that acute exposure to low concentration of DEHP does not compromise the function of HGrC1 cells, whereas high concentration causes only subtle effects. The identified nine novel targets of high-dose DEHP require further investigation to determine their role and importance in DEHP-exposed human granulosa cells.
邻苯二甲酸二（2-乙基己基）酯（DEHP）是一种内分泌干扰物，它对人颗粒细胞具有抗甾体生成作用；然而，这种作用的程度取决于 DEHP 的浓度和用于暴露的颗粒细胞模型。本研究的目的是确定低剂量和高剂量 DEHP 暴露对人颗粒细胞的影响。我们将人颗粒细胞系 HGrC1 暴露于 3 nM 和 25 μM 的 DEHP 中 48 小时。使用 DNBSEQ 测序平台和生物信息学工具分析全基因组转录组。结果表明，3 nM 的 DEHP 不会影响全局基因表达，而 25 μM 的 DEHP 仅影响 HGrC1 细胞中 9 个基因的表达：ABCA1、SREBF1、MYLIP、TUBB3、CENPT、NUPR1、ASS1、PCK2 和 CTSD。我们证实了 ABCA1mRNA 和 SREBP-1 蛋白（由 SREBF1 基因编码）的下调，这两者都参与胆固醇稳态。尽管存在这些变化，但在低剂量和高剂量 DEHP 暴露的 HGrC1 细胞中，孕激素的产生仍不受影响。高浓度的 DEHP 降低了 ABC1A mRNA 和 SREBP-1 蛋白的水平，并阻止了 forskolin 刺激的 HGrC1 细胞中孕激素合成蛋白 STAR 的上调；然而，观察到的变化不足以改变 forskolin 刺激的 HGrC1 细胞中孕激素的产生。总的来说，这项研究表明，急性暴露于低浓度的 DEHP 不会损害 HGrC1 细胞的功能，而高浓度只会引起轻微的影响。高剂量 DEHP 确定的 9 个新靶标需要进一步研究，以确定它们在暴露于 DEHP 的人颗粒细胞中的作用和重要性。
