Zhang Xuelian, Zhang Mengyang, Wang Chunzhi, Song Qingchuan, Yang Haiyan, Tang Qi, Zhao Qiaoling, Wang Jing, Pan Chuanying
Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China.
Jiangsu Key Laboratory of Sericultural Biology and Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, China.
Cells. 2025 Jun 20;14(13):940. doi: 10.3390/cells14130940.
Chlorpyrifos (CPF) has been extensively utilized in recent decades due to its highly efficient insecticidal properties. However, the widespread use of pesticides has posed new challenges to male reproduction. This study aims to explore the potential molecular mechanisms of male reproductive decline induced by CPF. We employ flow cytometry, qRT-PCR, Western blot, RNA sequencing, and bioinformatics analysis to investigate the potential molecular mechanisms involved in CPF-induced male reproductive damage in GC2 cells. Our results revealed that after 24 h of CPF treatment, the cell viability, cell cycle, apoptosis, and reactive oxygen species (ROS) accumulation of GC2 cells were significantly affected in vitro. RNA sequencing analysis data indicated that a total of 626 genes were differentially expressed compared to the DMSO group, especially for , , and . These differential genes were mainly enriched in signaling pathways such as PI3K-AKT and glutamine metabolism. In addition, further validation through qRT-PCR, Western blot, and experiments involving the inhibition of intracellular ROS generation with N-acetylcysteine collectively confirmed that CPF induces male reproductive damage through the ROS/AKT/Efcab6 pathway. These studies elucidate potential targets and molecular mechanisms underlying CPF-induced male infertility, providing a theoretical basis for the prevention of male reproductive damage caused by pesticide residues.
近几十年来，毒死蜱（CPF）因其高效的杀虫特性而被广泛使用。然而，农药的广泛使用给男性生殖带来了新的挑战。本研究旨在探讨CPF诱导男性生殖功能衰退的潜在分子机制。我们采用流式细胞术、qRT-PCR、蛋白质免疫印迹法、RNA测序和生物信息学分析，来研究CPF诱导GC2细胞雄性生殖损伤所涉及的潜在分子机制。我们的结果显示，在CPF处理24小时后，体外GC2细胞的细胞活力、细胞周期、凋亡和活性氧（ROS）积累均受到显著影响。RNA测序分析数据表明，与二甲基亚砜（DMSO）组相比，共有626个基因存在差异表达，尤其是[此处原文缺失具体基因名称]、[此处原文缺失具体基因名称]和[此处原文缺失具体基因名称]。这些差异基因主要富集在PI3K-AKT和谷氨酰胺代谢等信号通路中。此外，通过qRT-PCR、蛋白质免疫印迹法以及用N-乙酰半胱氨酸抑制细胞内ROS生成的实验进一步验证，共同证实CPF通过ROS/AKT/Efcab6途径诱导雄性生殖损伤。这些研究阐明了CPF诱导男性不育的潜在靶点和分子机制，为预防农药残留导致的男性生殖损伤提供了理论依据。
