Najafi Abouzar, Sharifi Seyed Davood, Farhadi Ramin, Bapir Shorsh Hussein
Department of Animal and Poultry Science, Faculty of Agricultural Technology, College of Agriculture and Natural Resources, University of Tehran, Tehran, Iran.
Department of Animal Science, Faculty of Agriculture, University of Kurdistan, Sanandaj, Iran.
Sci Rep. 2025 Jul 2;15(1):23564. doi: 10.1038/s41598-025-09943-7.
Cryopreservation of rooster sperm is a vital technique in avian reproductive management; however, it is often hindered by oxidative stress induced by reactive oxygen species (ROS) that negatively impact sperm quality during the freezing-thawing process. The present study aimed to investigate the impact of elamipretide, a mitochondria-targeted peptide, on sperm quality post-thaw. Sperm samples from 32-week-old broiler breeder roosters were cryopreserved using a Lake extender buffer with glycerol as the cryoprotectant. Four different concentrations of elamipretide (0, 6, 9, and 12 µmol/L) were tested in combination with the extender. Post-thaw sperm quality was evaluated by assessing motility, kinematic parameters, membrane integrity, mitochondrial activity, ROS levels as a direct marker of oxidative stress, lipid peroxidation, cell viability, and antioxidant enzyme activities (SOD, GPx and TAC). Sperm motility increased significantly at the 6 µmol/L (60.33 ± 1.54) and 9 µmol/L (64.96 ± 1.96) concentrations compared to the control, with the highest straight-line velocity observed at 9 µmol/L (21.21 ± 0.59). Membrane integrity also improved significantly at 9 µmol/L (61.78 ± 2.70) compared to lower doses (36.30 ± 1.64) and decreased at 12 µmol/L (49.57 ± 1.63). ROS production was significantly lower at 6 µmol/L (2.88 ± 0.07). Mitochondrial activity peaked at 9 µmol/L (60.21 ± 1.92), reflecting enhanced cell vitality and function. However, the effects were diminished at 12 µmol/L, indicating toxicity at higher concentrations. This study demonstrates the potential of elamipretide to improve rooster sperm cryopreservation outcomes by mitigating oxidative damage and preserving sperm quality post-thaw.
公鸡精子的冷冻保存是禽类繁殖管理中的一项重要技术；然而，它常常受到活性氧（ROS）诱导的氧化应激的阻碍，这种氧化应激在冻融过程中会对精子质量产生负面影响。本研究旨在探讨线粒体靶向肽依拉米肽对解冻后精子质量的影响。使用含有甘油作为冷冻保护剂的莱克稀释液对32周龄肉种鸡的精子样本进行冷冻保存。将四种不同浓度的依拉米肽（0、6、9和12 μmol/L）与稀释液联合测试。通过评估活力、运动学参数、膜完整性、线粒体活性、作为氧化应激直接标志物的ROS水平、脂质过氧化、细胞活力和抗氧化酶活性（超氧化物歧化酶、谷胱甘肽过氧化物酶和总抗氧化能力）来评估解冻后精子质量。与对照组相比，6 μmol/L（60.33±1.54）和9 μmol/L（64.96±1.96）浓度下精子活力显著提高，9 μmol/L时观察到最高直线速度（21.21±0.59）。与较低剂量（36.30±1.64）相比，9 μmol/L时膜完整性也显著改善（61.78±2.70），而在12 μmol/L时降低（49.57±1.63）。6 μmol/L时ROS产生显著降低（2.88±0.07）。线粒体活性在9 μmol/L时达到峰值（60.21±1.92），反映出细胞活力和功能增强。然而，在12 μmol/L时效果减弱，表明高浓度时具有毒性。本研究证明了依拉米肽通过减轻氧化损伤和保存解冻后精子质量来改善公鸡精子冷冻保存结果的潜力。
