背景:邻苯二甲酸盐暴露是一个主要的公共卫生问题,由于在环境中积累,并且与睾酮水平降低有关,导致不良妊娠结局。然而,邻苯二甲酸酯诱导的睾酮水平下降与铁凋亡之间的关系尚不明确.
目的:在此,我们旨在探讨邻苯二甲酸盐诱导的睾酮合成障碍的机制及其与铁凋亡的关系。
方法:我们在体内雄性小鼠模型和体外小鼠LeydigTM3细胞系中进行了验证实验,然后进行RNA测序和代谢组学分析。我们通过使用qRT-PCR和Western印迹评估了睾酮合成相关酶和铁凋亡相关指标的水平。然后,我们分析了脂质过氧化,ROS,Fe2+程度和谷胱甘肽体系证实了铁凋亡的发生。
结果:在本研究中,我们使用邻苯二甲酸二(2-乙基己基)酯(DEHP)来确定铁死亡是邻苯二甲酸酯诱导的睾酮水平下降的关键因素.已证明DEHP在Leydig细胞中引起谷胱甘肽代谢和类固醇合成障碍。作为DEHP的主要代谢产物,邻苯二甲酸单-2-乙基己酯(MEHP)引发的睾丸激素合成障碍伴随着溶质载体家族7成员11(SLC7A11)蛋白表达的减少。此外,MEHP通过增加细胞内和线粒体ROS与Erastin协同诱导铁凋亡,和脂质过氧化的产生。机械上,SLC7A11的过表达抵消了MEHP-Erastin共同暴露的协同作用。
结论:我们的研究结果表明,MEHP不会诱导铁凋亡,但会协同伊拉斯汀诱导的铁凋亡。这些发现为铁死亡在邻苯二甲酸酯诱导的睾丸激素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标。本研究建立了铁中毒与邻苯二甲酸酯细胞毒性之间的相关性,为减轻男性生殖疾病和“全球塑料毒性债务”的问题提供了一个新的观点。
BACKGROUND: 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.
OBJECTIVE: Herein, we aimed to explore the mechanisms of phthalates-induced testosterone synthesis disorder and its relationship to ferroptosis.
METHODS: 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, Fe2+ levels and glutathione system to confirm the occurrence of ferroptosis.
RESULTS: 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.
CONCLUSIONS: 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\".