Abstract：
Glyphosate (GLY), a widely used herbicide, can adversely affect the male reproductive health by inhibiting testosterone synthesis. Ferroptosis is a form of iron-dependent oxidative cell death that contributes to inhibition of testosterone secretion. However, it still remains unclear whether ferroptosis is involved in GLY-inhibited testosterone synthesis. Hereby, an in vitro model of 1 mM GLY-exposed testicular Leydig (TM3) cells was established to elucidate this issue. Data firstly showed that GLY causes cytotoxicity and testosterone synthesis inhibition via ferroptosis, while accumulation of lipid peroxides due to intracellular ferrous ion (Fe2+) overload and glutathione depletion is confirmed as a determinant of ferroptosis. Blockage of ferroptosis via chelation of Fe2+ or inhibition of lipid peroxidation can markedly mitigate GLY-induced testosterone synthesis inhibition. Also, autophagy activation is revealed in GLY-treated TM3 cells and nuclear receptor coactivator 4 (NCOA4)-mediated ferritinophagy is involved in ferroptosis through the release of excess Fe2+. GLY-induced cytotoxicity and testosterone synthesis inhibition are significantly alleviated by NCOA4 knockdown, demonstrating the crucial role of NCOA4-mediated ferritinophagy in GLY-inhibited testosterone synthesis. In summary, this study provides solid evidence that NCOA4-mediated ferritinophagy promotes ferroptosis to inhibit testosterone synthesis, highlighting that targeting NCOA4 may be a potential therapeutic approach in GLY-induced male reproductive toxicity.
摘要：
草甘膦(GLY),一种广泛使用的除草剂，可以通过抑制睾酮合成对男性生殖健康产生不利影响。铁凋亡是铁依赖性氧化细胞死亡的一种形式，有助于抑制睾酮分泌。然而,目前尚不清楚铁凋亡是否参与GLY抑制的睾酮合成。特此,建立了1mMGLY暴露的睾丸Leydig（TM3）细胞的体外模型来阐明这一问题。数据首先表明GLY通过铁凋亡引起细胞毒性和睾酮合成抑制，而由于细胞内亚铁离子（Fe2）过载和谷胱甘肽消耗而导致的脂质过氧化物的积累被证实是铁死亡的决定因素。通过螯合Fe2+或抑制脂质过氧化来阻断铁凋亡可显著减轻GLY诱导的睾酮合成抑制。此外，在GLY处理的TM3细胞中发现自噬激活，核受体共激活因子4（NCOA4）介导的铁细胞通过释放过量的Fe2参与铁细胞凋亡。GLY诱导的细胞毒性和睾酮合成抑制被NCOA4敲低显著缓解，证明NCOA4介导的铁素吞噬在GLY抑制的睾酮合成中的关键作用。总之,这项研究提供了确凿的证据，证明NCOA4介导的铁细胞吞噬促进铁细胞凋亡以抑制睾酮合成，强调靶向NCOA4可能是GLY诱导的男性生殖毒性的潜在治疗方法。
