Zhao Yi, Wang Xue-Qi, Liu Rui-Qi, Jiang Fu-Wei, Wang Jia-Xin, Chen Ming-Shan, Zhang Hao, Cui Jia-Gen, Chang Yuan-Hang, Li Jin-Long
College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, PR China.
College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, PR China; Key Laboratory of the Provincial Education Department of Heilongjiang for Common Animal Disease Prevention and Treatment, Northeast Agricultural University, Harbin 150030, PR China; Heilongjiang Key Laboratory for Laboratory Animals and Comparative Medicine, Northeast Agricultural University, Harbin 150030, PR China.
J Adv Res. 2025 May;71:369-381. doi: 10.1016/j.jare.2024.05.026. Epub 2024 May 24.
Phthalates exposure is a major public health concern due to the accumulation in the environment and associated with levels of testosterone reduction, leading to adverse pregnancy outcomes. However, the relationship between phthalate-induced testosterone level decline and ferroptosis remains poorly defined.
Herein, we aimed to explore the mechanisms of phthalates-induced testosterone synthesis disorder and its relationship to ferroptosis.
We conducted validated experiments in vivo male mice model and in vitro mouse Leydig TM3 cell line, followed by RNA sequencing and metabolomic analysis. We evaluated the levels of testosterone synthesis-associated enzymes and ferroptosis-related indicators by using qRT-PCR and Western blotting. Then, we analyzed the lipid peroxidation, ROS, Fe levels and glutathione system to confirm the occurrence of ferroptosis.
In the present study, we used di (2-ethylhexyl) phthalate (DEHP) to identify ferroptosis as the critical contributor to phthalate-induced testosterone level decline. It was demonstrated that DEHP caused glutathione metabolism and steroid synthesis disorders in Leydig cells. As the primary metabolite of DEHP, mono-2-ethylhexyl phthalate (MEHP) triggered testosterone synthesis disorder accompanied by a decrease in the expression of solute carri1er family 7 member 11 (SLC7A11) protein. Furthermore, MEHP synergistically induced ferroptosis with Erastin through the increase of intracellular and mitochondrial ROS, and lipid peroxidation production. Mechanistically, overexpression of SLC7A11 counteracts the synergistic effect of co-exposure to MEHP-Erastin.
Our research results suggest that MEHP does not induce ferroptosis but synergizes Erastin-induced ferroptosis. These findings provide evidence for the role of ferroptosis in phthalates-induced testosterone synthesis disorder and point to SLC7A11 as a potential target for male reproductive diseases. This study established a correlation between ferroptosis and phthalates cytotoxicity, providing a novel view point for mitigating the issue of male reproductive disease and "The Global Plastic Toxicity Debt".
邻苯二甲酸盐暴露是一个重大的公共卫生问题，因为其会在环境中积累，并与睾酮水平降低相关，从而导致不良妊娠结局。然而，邻苯二甲酸盐诱导的睾酮水平下降与铁死亡之间的关系仍不清楚。
在此，我们旨在探讨邻苯二甲酸盐诱导睾酮合成障碍的机制及其与铁死亡的关系。
我们在体内雄性小鼠模型和体外小鼠睾丸间质细胞系TM3中进行了验证实验，随后进行RNA测序和代谢组学分析。我们使用qRT-PCR和蛋白质免疫印迹法评估睾酮合成相关酶和铁死亡相关指标的水平。然后，我们分析脂质过氧化、活性氧、铁水平和谷胱甘肽系统以确认铁死亡的发生。
在本研究中，我们使用邻苯二甲酸二（2-乙基己基）酯（DEHP）确定铁死亡是邻苯二甲酸盐诱导睾酮水平下降的关键因素。结果表明，DEHP导致睾丸间质细胞中谷胱甘肽代谢和类固醇合成紊乱。作为DEHP的主要代谢产物，邻苯二甲酸单-2-乙基己酯（MEHP）引发睾酮合成障碍，同时溶质载体家族7成员11（SLC7A11）蛋白表达下降。此外，MEHP通过增加细胞内和线粒体活性氧以及脂质过氧化产物，与埃拉斯汀协同诱导铁死亡。从机制上讲，SLC7A11的过表达抵消了共同暴露于MEHP-埃拉斯汀的协同作用。
我们的研究结果表明，MEHP不会诱导铁死亡，而是协同埃拉斯汀诱导铁死亡。这些发现为铁死亡在邻苯二甲酸盐诱导的睾酮合成障碍中的作用提供了证据，并指出SLC7A11是男性生殖疾病的潜在靶点。本研究建立了铁死亡与邻苯二甲酸盐细胞毒性之间的关联，为缓解男性生殖疾病问题和“全球塑料毒性债务”提供了新的视角。
