Fang Zishui, Jin Zirun, Zhao Qiancheng, Weng Jiaming, Zhang Zhe, Yang Yuzhuo, Jiang Hui
Department of Urology, Peking University First Hospital, Beijing 100034, China; Institution of Urology, Peking University, Beijing 100034, China; Beijing Key Laboratory of Urogenital Diseases (Male) Molecular Diagnosis and Treatment Center, Beijing 100034, China.
Department of Urology, Center for Reproductive Medicine, Peking University Third Hospital, No. 49 North Garden Road, Haidian District, Beijing 100191, China.
Reprod Toxicol. 2025 Mar;132:108758. doi: 10.1016/j.reprotox.2024.108758. Epub 2024 Nov 28.
Di-(2-ethylhexyl) phthalate (DEHP) exposure has been associated with male reproductive damage, but the mechanisms involved remain incompletely defined. This study aims to investigate the effects of DEHP exposure on the testes of prepubertal rats through an integrative analysis of metabolomics and transcriptomics, combined with molecular experiments. DEHP exposure resulted in decreased testis weight and increased oxidative stress level in the testis tissues of prepubertal male rats. Moreover, our findings showed a disordered testis structure, reduced spermatogenic and Sertoli cells as well as destruction of mitochondria structure in the testis tissues of DEHP-treated prepubertal male rats. Transcriptome function analysis together with metabolome function analysis indicated that spermatogenesis, apoptosis, inflammatory, lipid metabolism as well as DNA repair signaling pathway were enriched in the testis of DEHP-treated prepubertal male rats. The integrative omics analysis further suggested that TNF-α induced apoptosis played a crucial role in mediating the detrimental effects of DEHP exposure on the testis of prepubertal rats, which was validated by ELISA, Western blotting and Tunel assays. Validation experiments conducted in vitro using GC-2 cells corroborated these findings, demonstrating that mono-(2-ethylhexyl) phthalate (MEHP), the main active metabolite of DEHP, significantly inhibits cell proliferation and increases apoptosis via activating the TNF-α apoptosis pathway. Overall, these findings provided a novel mechanism of dysregulated spermatogenesis of DEHP exposure on the testes of prepubertal rats.
邻苯二甲酸二（2-乙基己基）酯（DEHP）暴露与雄性生殖损伤有关，但其涉及的机制仍未完全明确。本研究旨在通过代谢组学和转录组学的综合分析，并结合分子实验，探讨DEHP暴露对青春期前大鼠睾丸的影响。DEHP暴露导致青春期前雄性大鼠睾丸组织重量减轻，氧化应激水平升高。此外，我们的研究结果显示，DEHP处理的青春期前雄性大鼠睾丸组织结构紊乱，生精细胞和支持细胞减少，线粒体结构破坏。转录组功能分析和代谢组功能分析表明，在DEHP处理的青春期前雄性大鼠睾丸中，精子发生、细胞凋亡、炎症、脂质代谢以及DNA修复信号通路均有富集。综合组学分析进一步表明，肿瘤坏死因子-α（TNF-α）诱导的细胞凋亡在介导DEHP暴露对青春期前大鼠睾丸的有害影响中起关键作用，这通过酶联免疫吸附测定（ELISA）、蛋白质免疫印迹法（Western blotting）和脱氧核糖核苷酸末端转移酶介导的缺口末端标记法（Tunel）检测得到验证。使用GC-2细胞进行的体外验证实验证实了这些发现，表明DEHP的主要活性代谢物单（2-乙基己基）邻苯二甲酸酯（MEHP）通过激活TNF-α凋亡途径显著抑制细胞增殖并增加细胞凋亡。总体而言，这些发现为DEHP暴露导致青春期前大鼠睾丸精子发生失调提供了一种新机制。
