Guo Lewei, Gu Zhuoxuan, Wang Bing, Wang Yunuo, Chen Jiaorong, Li Yitong, Zheng Qiuju, Zhao Jing, Ding He, Liu Hongyu, Fang Yi, Wang Jun, Lyu Wenfa
Key Laboratory of Animal Production, Product Quality and Security, Ministry of Education, Jilin Agricultural University, Changchun 130118, China.
Key Laboratory of Utilization and Protection of Beef Cattle Germplasm Resources, Jilin Agricultural University, Changchun 130118, China.
Antioxidants (Basel). 2025 Jul 14;14(7):861. doi: 10.3390/antiox14070861.
Sperm freezability exhibits marked individual variability, yet the mechanisms remain unclear. Using bulls as the experimental model, we integrated proteomic (sperm) and metabolomic (seminal plasma) analyses of high-freezability (HF) and control (CF) bulls to identify key biomarkers associated with sperm freezability. Post-thaw motility and membrane integrity were significantly higher in HF bulls ( < 0.05). Sperm proteome analysis revealed upregulated antioxidant proteins (PRDX2, GSTM4), heat shock proteins (HSP70, HSP90), and key enzymes in arginine and proline metabolism (PRODH, LAP3). Seminal plasma metabolomics revealed elevated spermine in HF bulls. Meanwhile, we found that spermine abundance was positively correlated with post-thaw motility, as well as with the expression levels of both PRODH and LAP3 (r > 0.6, < 0.05). Functional validation demonstrated that 200 μM spermine supplementation in cryopreservation extenders enhanced post-thaw motility, kinematic parameters (VAP, VSL, VCL), membrane integrity, and acrosome integrity ( < 0.05). Concurrently, spermine enhanced antioxidant enzyme (SOD, CAT, GSH-Px) activity and reduced ROS and MDA levels ( < 0.05). Our study reveals a spermine-driven antioxidant network coordinating sperm-seminal plasma synergy during cryopreservation, offering novel strategies for semen freezing optimization.
精子的冷冻保存能力存在显著的个体差异，但其机制仍不清楚。我们以公牛为实验模型，整合了高冷冻保存能力（HF）公牛和对照（CF）公牛的蛋白质组学（精子）和代谢组学（精浆）分析，以确定与精子冷冻保存能力相关的关键生物标志物。解冻后，HF公牛的精子活力和膜完整性显著更高（<0.05）。精子蛋白质组分析显示，抗氧化蛋白（PRDX2、GSTM4）、热休克蛋白（HSP70、HSP90）以及精氨酸和脯氨酸代谢中的关键酶（PRODH、LAP3）上调。精浆代谢组学显示，HF公牛的精胺水平升高。同时，我们发现精胺丰度与解冻后精子活力以及PRODH和LAP3的表达水平呈正相关（r>0.6，<0.05）。功能验证表明，在冷冻保存稀释液中添加200μM精胺可提高解冻后精子活力、运动学参数（VAP、VSL、VCL）、膜完整性和顶体完整性（<0.05）。同时，精胺增强了抗氧化酶（SOD、CAT、GSH-Px）的活性，降低了ROS和MDA水平（<0.05）。我们的研究揭示了一个由精胺驱动的抗氧化网络，该网络在冷冻保存过程中协调精子与精浆的协同作用，为优化精液冷冻提供了新策略。
