Germline Stem Cells and Microenvironment Lab, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China; Stem Cell Research and Translation Center, Nanjing Agricultural University, Nanjing 210095, China.
Germline Stem Cells and Microenvironment Lab, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China.
Reprod Biol. 2023 Sep;23(3):100781. doi: 10.1016/j.repbio.2023.100781. Epub 2023 Jun 6.
Zearalenone (ZEA) is a prevalent mycotoxin found in moldy diets and is associated with reproductive dysfunction. However, the molecular underpinning of ZEA in impairment of spermatogenesis remains largely unknown. To unveil the toxic mechanism of ZEA, we established a co-culture model using porcine Sertoli cells and porcine spermatogonial stem cells (pSSCs) to investigate the impact of ZEA on these cell types and their associated signaling pathways. Our findings showed that low concentration of ZEA inhibited cell apoptosis, while high concentration induced cell apoptosis. Furthermore, the expression levels of Wilms' tumor 1 (WT1), proliferating cell nuclear antigen (PCNA) and glial cell line-derived neurotrophic factor (GDNF) were significantly decreased in ZEA treatment group, while concurrently upregulating the transcriptional levels of the NOTCH signaling pathway target genes HES1 and HEY1. The addition of the NOTCH signaling pathway inhibitor DAPT (GSI-IX) alleviated the damage to porcine Sertoli cells caused by ZEA. Gastrodin (GAS) significantly increased the expression levels of WT1, PCNA and GDNF, and inhibited the transcription of HES1 and HEY1. GAS also efficiently restored the decreased expression levels of DDX4, PCNA and PGP9.5 in co-cultured pSSCs suggesting its potential in ameliorating the damage caused by ZEA to Sertoli cells and pSSCs. In conclusion, the present study demonstrates that ZEA disrupts pSSCs self-renewal by affecting the function of porcine Sertoli cell, and highlights the protective mechanism of GAS through the regulation of the NOTCH signaling pathway. These findings may offer a novel strategy for alleviating ZEA-induced male reproductive dysfunction in animal production.
玉米赤霉烯酮（ZEA）是一种常见的霉菌毒素，存在于霉变的饲料中，与生殖功能障碍有关。然而，ZEA 损害精子发生的分子基础在很大程度上仍不清楚。为了揭示 ZEA 的毒性机制，我们建立了一个猪睾丸支持细胞和猪精原干细胞（pSSC）的共培养模型，以研究 ZEA 对这些细胞类型及其相关信号通路的影响。我们的研究结果表明，低浓度的 ZEA 抑制细胞凋亡，而高浓度的 ZEA 诱导细胞凋亡。此外，在 ZEA 处理组中，Wilms 肿瘤 1（WT1）、增殖细胞核抗原（PCNA）和胶质细胞源性神经营养因子（GDNF）的表达水平显著降低，同时NOTCH 信号通路靶基因 HES1 和 HEY1 的转录水平上调。添加 NOTCH 信号通路抑制剂 DAPT（GSI-IX）可减轻 ZEA 对猪睾丸支持细胞的损伤。天麻素（GAS）显著增加 WT1、PCNA 和 GDNF 的表达水平，并抑制 HES1 和 HEY1 的转录。GAS 还有效地恢复了共培养的 pSSC 中 DDX4、PCNA 和 PGP9.5 的表达水平降低，表明其在改善 ZEA 对睾丸支持细胞和 pSSC 损伤方面的潜力。总之，本研究表明，ZEA 通过影响猪睾丸支持细胞的功能破坏 pSSC 的自我更新，并强调了 GAS 通过调节 NOTCH 信号通路的保护机制。这些发现可能为缓解动物生产中 ZEA 诱导的雄性生殖功能障碍提供新策略。
