Liang Yongchao, Huo Hongjin, Wei Wei, Wang Dongxuan, Lu Jingjing, Yi Wenjie, Jiang Fei
Department of Occupational and Environmental Health, School of Public Health, Suzhou Medical College of Soochow University, Suzhou, China.
Jiangsu Key Laboratory of Preventive and Translational Medicine for Geriatric Diseases, MOE Key Laboratory of Geriatric Diseases and Immunology, Suzhou Medical College of Soochow University, Suzhou, China.
J Endocrinol Invest. 2025 Jul 19. doi: 10.1007/s40618-025-02656-8.
Perfluorooctane sulfonic acid (PFOS) is a representative persistent organic pollutant that has been shown to impair male reproductive function. Epidemiological studies and animal models suggest that vitamin D may modify male reproductive function. However, whether 1α,25(OH)D could mitigate the PFOS-induced damage of Sertoli cells, and the underlying mechanisms remain poorly defined.
Mouse Sertoli cell line TM4 were pretreated with or without 10 nM or 100 nM 1α,25(OH)D for 6 h, followed by 100 µM or 200 µM PFOS for 24 h. Cell viability was assessed using the CCK8 assay. Reactive oxygen species (ROS) were detected by fluorescent staining, while malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) levels were measured using commercial kits. siRNA transfection, quantitative real-time PCR (qRT‒PCR), and Western blotting were performed to elucidate the underlying mechanisms. Molecular docking studies were conducted to explore the potential binding of PFOS to Keap1.
PFOS increased MDA contents, decreased GSH levels, and induced cytotoxicity in mouse Sertoli cells. Mechanistically, PFOS might directly bind to Keap1, facilitating its degradation, leading to persistent activation of the Nrf2-ARE pathway. Conversely, 1α,25(OH)D attenuated PFOS-induced cytotoxicity by alleviating the Nrf2-ARE pathway, a protective effect which was abolished by vitamin D receptor (VDR) knockdown.
PFOS induces cytotoxicity in Sertoli cells by binding to Keap1 and persistently activating the Nrf2-ARE pathway. 1α,25(OH)D emerges as a promising candidate for mitigating PFOS-induced testicular dysfunction.
全氟辛烷磺酸（PFOS）是一种典型的持久性有机污染物，已被证明会损害雄性生殖功能。流行病学研究和动物模型表明，维生素D可能会改变雄性生殖功能。然而，1α,25(OH)D是否能减轻PFOS诱导的支持细胞损伤，其潜在机制仍不清楚。
小鼠支持细胞系TM4分别用10 nM或100 nM 1α,25(OH)D预处理6小时，然后用100 µM或200 µM PFOS处理24小时。使用CCK8法评估细胞活力。通过荧光染色检测活性氧（ROS），同时使用商业试剂盒测量丙二醛（MDA）、超氧化物歧化酶（SOD）和谷胱甘肽（GSH）水平。进行小干扰RNA（siRNA）转染、定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法以阐明潜在机制。进行分子对接研究以探索PFOS与Keap1的潜在结合。
PFOS增加了小鼠支持细胞中MDA含量，降低了GSH水平，并诱导了细胞毒性。机制上，PFOS可能直接与Keap1结合，促进其降解，导致Nrf2-ARE途径持续激活。相反，1α,25(OH)D通过减轻Nrf2-ARE途径减轻了PFOS诱导的细胞毒性，维生素D受体（VDR）敲低消除了这种保护作用。
PFOS通过与Keap1结合并持续激活Nrf2-ARE途径诱导支持细胞产生细胞毒性。1α,25(OH)D是减轻PFOS诱导的睾丸功能障碍的有希望的候选物。
