Li Yang, Liu Xue, Cheng Ye, Li Jingchun, Zhou Yuling, Guo Qing
College of Animal Science and Technology, Heilongjiang Bayi Agricultural University, Daqing, China.
Hangzhou Dimoman Biotechnology, Hangzhou, China.
Front Vet Sci. 2025 Jul 31;12:1611661. doi: 10.3389/fvets.2025.1611661. eCollection 2025.
The plasma membrane of boar sperm is notably enriched in polyunsaturated fatty acids (PUFAs). During extended liquid storage of boar semen at 17°C, reactive oxygen species (ROS) derived from lipid peroxidation progressively accumulate within sperm cells. Concurrently, the onset of ferroptosis is initiated by the disruption of intracellular redox homeostasis, characterized by an imbalance between the production and elimination of lipid-derived ROS. This study aims to investigate whether the ferroptosis inhibitor Liproxstatin-1 (Lip-1) protects boar sperm quality during 17°C liquid preservation by ameliorating oxidative stress and regulating ferroptosis markers.
Various concentrations of Lip-1 were added to the modified Modena extender, and sperm motility and kinetic parameters were assessed using the CASA system, which facilitated the identification of the optimal Lip-1 concentration. Subsequently, the integrity of the acrosome, plasma membrane, and mitochondrial membrane potential (MMP) of sperm was examined in both the control group and the optimal of Lip-1 group. Additionally, the antioxidant capacity and lipid peroxidation levels of the sperm were evaluated. Furthermore, the ferroptosis inducer Erastin (Era) was utilized to investigate whether Lip-1 could regulate oxidative stress and ferroptosis markers to enhance the liquid preservation efficiency of boar semen at 17°C.
Various concentrations of Lip-1 were added to the modified Modena extender, and the results indicated that, compared to the control group, 0.2 μM of Lip-1 significantly enhanced sperm motility and kinetic parameters. Additionally, a concentration of 0.2 μM Lip-1 significantly enhanced sperm quality, which included improvements in the integrity of the sperm plasma membrane and acrosome, antioxidant capacity, and MMP. Additional, additional tests revealed that Lip-1 can significantly reduce markers of sperm lipid peroxidation during the room temperature preservation of boar semen, including C11-bodipy, MDA, LPO, and improved ferroptosis-related protein GPX4. Furthermore, the ferroptosis inducer Era was utilized, and the results demonstrated that 0.2 μM Lip-1 significantly alleviated the sperm damage induced by Era.
The results of this study indicated that Lip-1 significantly enhanced the liquid preservation efficiency of boar semen at 17°C associated with ameliorating oxidative stress and regulating ferroptosis markers, providing both theoretical and practical references for improving the liquid preservation of boar semen.
公猪精子的质膜富含多不饱和脂肪酸（PUFAs）。在公猪精液于17°C进行长时间液体保存期间，脂质过氧化产生的活性氧（ROS）在精子细胞内逐渐积累。同时，细胞内氧化还原稳态的破坏引发铁死亡，其特征是脂质来源的ROS产生与清除之间的失衡。本研究旨在探讨铁死亡抑制剂Liproxstatin-1（Lip-1）是否通过减轻氧化应激和调节铁死亡标志物来保护17°C液体保存期间的公猪精子质量。
将不同浓度的Lip-1添加到改良的摩德纳稀释液中，使用计算机辅助精子分析（CASA）系统评估精子活力和运动参数，以确定最佳Lip-1浓度。随后，检测对照组和最佳Lip-1浓度组精子顶体、质膜的完整性以及线粒体膜电位（MMP）。此外，评估精子的抗氧化能力和脂质过氧化水平。此外，利用铁死亡诱导剂Erastin（Era）研究Lip-1是否能调节氧化应激和铁死亡标志物，以提高公猪精液在17°C下的液体保存效率。
将不同浓度的Lip-1添加到改良的摩德纳稀释液中，结果表明，与对照组相比，0.2μM的Lip-1显著提高了精子活力和运动参数。此外，0.2μM Lip-1浓度显著提高了精子质量，包括精子质膜和顶体完整性、抗氧化能力以及MMP的改善。此外，进一步测试表明，Lip-1可在公猪精液室温保存期间显著降低精子脂质过氧化标志物，包括C11-硼二吡咯、丙二醛（MDA）、脂质过氧化物（LPO），并改善铁死亡相关蛋白谷胱甘肽过氧化物酶4（GPX4）。此外，利用铁死亡诱导剂Era，结果表明0.2μM Lip-1显著减轻了Era诱导的精子损伤。
本研究结果表明，Lip-1显著提高了公猪精液在17°C下的液体保存效率，这与减轻氧化应激和调节铁死亡标志物有关，为提高公猪精液的液体保存提供了理论和实践参考。
