Bis(2-ethylhexyl)tetrabromophthalate (TBPH) has been widely detected in the environment and organisms; thus, its toxic effects on male reproduction were systematically studied. First, we found that TBPH can stably bind to the androgen receptor (AR) based on in silico molecular docking results and observed an antagonistic activity, but not agonistic activity, on the AR signaling pathway using a constructed AR-GRIP1 yeast assay. Subsequently, we validated the adverse effects on male germ cells by observing inhibited androgen production and proliferation in Leydig cells upon in vitro exposure and affected general motility and motive tracks of zebrafish sperm upon ex vivo exposure. Finally, the in vivo reproductive toxicity was demonstrated in male zebrafish by reduced mating behavior in F0 generation when paired with unexposed females and abnormal development of their offspring. In addition, reduced sperm motility and impaired germ cells in male zebrafish were also observed, which may be related to the disturbed homeostasis of sex hormones. Notably, the specifically suppressed AR in the brain provides further evidence for the antagonistic effects as above-mentioned. These results confirmed that TBPH affected male reproduction through a classical nuclear receptor-mediated pathway, which would be helpful for assessing the ecological and health risks of TBPH.

中文翻译：

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四溴邻苯二甲酸二（2-乙基己基）酯男性生殖毒性的综合研究：计算机、体外、离体和体内

四溴邻苯二甲酸二（2-乙基己基）酯（TBPH）已在环境和生物体中广泛检测到；因此，系统地研究了它对男性生殖的毒性作用。首先，基于计算机分子对接结果，我们发现 TBPH 可以稳定地与雄激素受体 (AR) 结合，并使用构建的 AR-GRIP1 酵母分析观察到对 AR 信号通路的拮抗活性，但没有激动活性。随后，我们通过观察体外暴露时睾丸间质细胞雄激素产生和增殖受到抑制以及体外暴露时斑马鱼精子的一般运动和运动轨迹的影响来验证对雄性生殖细胞的不利影响。最后，雄性斑马鱼与未暴露的雌性斑马鱼配对时，F0代的交配行为减少，并且其后代发育异常，从而证明了雄性斑马鱼的体内生殖毒性。此外，还观察到雄性斑马鱼的精子活力降低和生殖细胞受损，这可能与性激素稳态紊乱有关。值得注意的是，大脑中特异性抑制的 AR 为上述拮抗作用提供了进一步的证据。这些结果证实TBPH通过经典的核受体介导途径影响男性生殖，这有助于评估TBPH的生态和健康风险。