Selenium is a necessary trace element for animals, and it can improve the antioxidant capacity of a variety of cells. In this study, the antioxidant capacity of different concentrations of selenium nanoparticles (SeNPs) (0.1, 0.5, 1.5, and 4.5 mg/L) on Onychostoma macrolepis and Schizothorax prenanti sperm were explored during stored at 4 °C for 72 h. After 24 h, 0.5 mg/L SeNPs was more effective at improving the quality of Onychostoma macrolepis sperm by stabilizing the level of reactive oxygen species (ROS) within the sperm. A similar function of SeNPs was also found in Schizothorax prenanti sperm. RAS-selective lethal 3 (RSL3) significantly inhibited the effect of SeNPs on Onychostoma macrolepis sperm quality. Moreover, RSL3 completely abolished the effect of SeNPs which inhibited glutathione peroxidase 4 (GPX4) protein degradation in Onychostoma macrolepis sperm. The GPX4 protein is mainly located at the midpiece, where mitochondria were enriched in Onychostoma macrolepis sperm. These results revealed that GPX4 played an important role in sperm storage in vitro, and SeNPs stabilized ROS levels mainly by regulating GPX4 in Onychostoma macrolepis sperm. Furthermore, compared with the midpiece of the non-activated sperm, the midpiece of the activated sperm displayed deformation and a higher ATP content. In summary, SeNPs can effectively sustain the level of physiological reactive oxygen species (ROS) in Onychostoma macrolepis and Schizothorax prenanti sperm during storage at 4 °C, thereby SeNPs may be a potential additive for fish sperm stored in vitro.

硒是动物必需的微量元素，能提高多种细胞的抗氧化能力。在本研究中，研究了不同浓度的硒纳米颗粒（SeNPs）（0.1、0.5、1.5 和 4.5 mg/L）在 4 °C 下储存 72 h 期间对大鳞甲线虫和Schizothorax prenanti精子的抗氧化能力。24 小时后，0.5 mg/L SeNPs通过稳定精子内的活性氧 (ROS) 水平，更有效地提高了大鳞甲鱼精子的质量。在Schizothorax prenanti精子中也发现了类似的 SeNPs 功能。RAS-selective lethal 3 (RSL3) 显着抑制 SeNPs 对Onychostoma macrolepis的影响精子质量。此外，RSL3 完全消除了 SeNPs 的作用，即抑制大鳞甲虫精子中谷胱甘肽过氧化物酶 4 (GPX4) 蛋白降解。GPX4蛋白主要位于中段，线粒体富含大鳞甲线虫精子。这些结果表明GPX4在体外精子储存中起重要作用，SeNPs主要通过调节大鳞甲鱼精子中的GPX4来稳定ROS水平。此外，与未活化精子的中段相比，活化精子的中段表现出变形和更高的ATP含量。总之，SeNPs 可以有效维持体内生理活性氧（ROS）的水平。Onychostoma macrolepis和Schizothorax prenanti精子在 4 °C 下储存期间，因此 SeNPs 可能是体外储存的鱼精子的潜在添加剂。