Department of Physiology, Faculty of Veterinary Medicine, Kirikkale University, 71450 Yahsihan, Kirikkale, Turkey; Deparment of Obstetrics and Gynecology, University of Nebraska Medical Center, Omaha, NE 68198, USA.
Deparment of Obstetrics and Gynecology, University of Nebraska Medical Center, Omaha, NE 68198, USA; Veterans Affairs Nebraska-Western Iowa Health Care System, Omaha, NE 68105, USA.
Environ Pollut. 2023 Dec 1;338:122698. doi: 10.1016/j.envpol.2023.122698. Epub 2023 Oct 11.
Perfluorooctanoic acid (PFOA) is a persistent environmental contaminant. Due to the ubiquitous presence of PFOA in the environment, the impacts of PFOA exposure not only affect human reproductive health but may also affect livestock reproductive health. The focus of this study was to determine the effects of PFOA on the physiological functions of bovine granulosa cells in vitro. Primary bovine granulosa cells were exposed to 0, 4, and 40 μM PFOA for 48 and 96 h followed by analysis of granulosa cell function including cell viability, steroidogenesis, and mitochondrial activity. Results revealed that PFOA inhibited steroid hormone secretion and altered the expression of key enzymes required for steroidogenesis. Gene expression analysis revealed decreases in mRNA transcripts for CYP11A1, HSD3B, and CYP19A1 and an increase in STAR expression after PFOA exposure. Similarly, PFOA decreased levels of CYP11A1 and CYP19A1 protein. PFOA did not impact live cell number, alter the cell cycle, or induce apoptosis, although it reduced metabolic activity, indicative of mitochondrial dysfunction. We observed that PFOA treatment caused a loss of mitochondrial membrane potential and increases in PINK protein expression, suggestive of mitophagy and mitochondrial damage. Further analysis revealed that these changes were associated with increased levels of reactive oxygen species. Expression of autophagy related proteins phosphoULK1 and LAMP2 were increased after PFOA exposure, in addition to an increased abundance of lysosomes, characteristic of increased autophagy. Taken together, these findings suggest that PFOA can negatively impact granulosa cell steroidogenesis via mitochondrial dysfunction.
全氟辛酸（PFOA）是一种持久性环境污染物。由于 PFOA 普遍存在于环境中，其暴露的影响不仅会影响人类生殖健康，还可能影响家畜生殖健康。本研究的重点是确定 PFOA 对体外牛颗粒细胞生理功能的影响。将原代牛颗粒细胞分别暴露于 0、4 和 40 μM PFOA 中 48 和 96 h，然后分析颗粒细胞功能，包括细胞活力、类固醇生成和线粒体活性。结果表明，PFOA 抑制类固醇激素的分泌，并改变类固醇生成所需的关键酶的表达。基因表达分析显示，暴露于 PFOA 后，CYP11A1、HSD3B 和 CYP19A1 的 mRNA 转录物减少，而 STAR 的表达增加。同样，PFOA 降低了 CYP11A1 和 CYP19A1 蛋白的水平。PFOA 不影响活细胞数量、改变细胞周期或诱导细胞凋亡，尽管它降低了代谢活性，表明存在线粒体功能障碍。我们观察到 PFOA 处理导致线粒体膜电位丧失和 PINK 蛋白表达增加，提示存在自噬和线粒体损伤。进一步的分析表明，这些变化与活性氧水平的增加有关。在暴露于 PFOA 后，自噬相关蛋白磷酸化 ULK1 和 LAMP2 的表达增加，同时溶酶体的丰度增加，表明自噬增加。总之，这些发现表明，PFOA 可通过线粒体功能障碍对颗粒细胞类固醇生成产生负面影响。
