Department of Biology and Ecology, Faculty of Sciences, University of Novi Sad, 21000 Novi Sad, Serbia.
Cells. 2022 Jul 27;11(15):2304. doi: 10.3390/cells11152304.
Here, we applied a model of long-term exposure of human granulosa cells to low environmentally relevant levels of di(2-ethylhexyl) phthalate (DEHP). This approach provides more relevant data regarding the impact of DEHP on the function of human granulosa cells. The immortalized human granulosa cells HGrC1 were exposed to 50 nM and 250 nM DEHP for four weeks. The cells were collected every week to analyze the basal granulosa cells' functions. A portion of the DEHP-exposed cells was stimulated with forskolin (FOR) for 48 h. Steroidogenesis was investigated using ELISA, whereas DNBQ sequencing and RT-qPCR were used to analyze gene expression. The results show that steroidogenesis was not affected by DEHP exposure. RNAsequencing shows that DEHP caused week- and concentration-specific changes in various genes and functions in HGrC1. Sulfotransferase family 1A member 3 () and 4 (), which are involved in catecholamine metabolism, were the most prominent genes affected by DEHP under both the basal and FOR-stimulated conditions in all four weeks of exposure. This study showed, for the first time, that and are expressed in human granulosa cells, are regulated by FOR, and are affected by low-level DEHP exposure. These data provide new insight into the relationship between DEHP, , and in human granulosa cells.
在这里，我们应用了一种人类颗粒细胞长期暴露于低环境相关浓度邻苯二甲酸二（2-乙基己基）酯（DEHP）的模型。这种方法提供了关于 DEHP 对人类颗粒细胞功能影响的更相关数据。永生化的人类颗粒细胞 HGrC1 被暴露于 50 nM 和 250 nM 的 DEHP 中四周。每周收集细胞以分析基础颗粒细胞的功能。一部分暴露于 DEHP 的细胞用 forskolin (FOR) 刺激 48 小时。使用 ELISA 研究类固醇生成，而使用 DNBQ 测序和 RT-qPCR 分析基因表达。结果表明，DEHP 暴露不影响类固醇生成。RNA 测序显示，DEHP 导致 HGrC1 中各种基因和功能在不同时间和浓度上出现特异性变化。磺基转移酶家族 1A 成员 3() 和 4()，参与儿茶酚胺代谢，是暴露于 DEHP 后在所有四周的基础和 FOR 刺激条件下受影响最显著的基因。这项研究首次表明，在人类颗粒细胞中表达，并受 FOR 调节，并且受到低水平 DEHP 暴露的影响。这些数据为 DEHP、和在人类颗粒细胞中的关系提供了新的见解。
