烟酸通过降低氧化应激水平提高绵羊精子低温保存效果

doi:10.11843/j.issn.0366-6964.2023.05.019

摘要/Abstract

摘要： 低温保存可有效延长精子体外保存时间,提高精液利用效率。但是长时间处于低温环境下会造成活性氧(reactive oxygen species,ROS)持续累积,诱发精子氧化应激,降低质膜完整性,造成精子活力下降。旨在研究绵羊精子4℃保存条件下,稀释液添加不同浓度烟酸(nicotinic acid,NA)对精子的保存效果,并探讨其作用机理。采集健康的成年德国美利奴种公羊精液(n=6),以基础稀释液中添加5、10、20 mmol·L^-1 NA为保护剂,拟阐明NA对绵羊精子的保护作用。通过精子分析仪检测精子活力、运动性能,精子低渗膨胀试验检测质膜完整性,荧光染色检测精子中ROS水平、线粒体功能,试剂盒检测H₂O₂含量、抗氧化因子活性水平及ATP水平。结果表明:与10%BSA对照组相比较,添加NA处理后精子的存活时间可延长至120 h,其中10 mmol·L^-1 NA NA处理后第72小时精子活力仍可达到70%以上;在NA处理后24~72 h时,绵羊精子可保持更高的活力、运动性能与及质膜完整性(P<0.05),而且降低了精子中的ROS水平(P<0.05)。在NA处理后第72小时,精子中H₂O₂含量显著低于10%BSA对照组(P<0.05),而精子中抗氧化因子(CAT、GSH-PX、T-SOD、T-AOC)活性水平、线粒体功能以及ATP水平均显著优于10%BSA对照组(P<0.05)。综上,在4℃保存条件下,绵羊精液低温稀释液中添加NA,可提高精子的活力、运动性能、质膜完整性和抗氧化能力,并改善精子线粒体功能和ATP生成,有效提升绵羊精子低温保存效果。

关键词: 绵羊, 烟酸, 精子活力, 线粒体, 氧化应激

Abstract: Low temperature preservation can effectively prolong the storage time of sperm in vitro and improve the semen utilization ratio. However, long-term exposure to low temperature will cause the continuous accumulation of reactive oxygen species (ROS), induce sperm oxidative stress, reduce the integrity of plasma membrane and result in sperm motility decline. This study intends to investigate the effect of adding different concentrations of niacin (nicotinic acid, NA) to the diluents on sperm preservation under the condition of 4℃ storage of sheep sperm, and to explore its mechanism of action. In this study, semen from healthy adult German Merino rams (n=6) were collected and supplemented with 5、10 and 20 mmol·L^-1 NA used as protective agents to clarify the protective effect of NA on sheep sperm. Sperm motility and motility were detected by sperm analyzer, plasma membrane integrity was analyzed by HOST,ROS level and mitochondrial function in sperm were detected by fluorescent staining, while H₂O₂ content, antioxidant factor activity level and ATP level were detected by kit. The results showed that compared with 10% BSA control group, the survival time of sperm treated with NA could be extended to 120 h,and the sperm motility could still reach 70% at 72 h after 10 mmol·L^-1NA treatment. At 24~72 h after NA treatment, sheep sperm maintained higher motility, movement characteristic and plasma membrane integrity (P<0.05), and decreased ROS levels in sperm cells (P<0.05). At 72 h after NA treatment, the content of H₂O₂in sperm was significantly lower than that in the 10% BSA control group (P<0.05), while the antioxidant factors (CAT,GSH-PX,T-SOD,T-AOC) activity levels, mitochondrial function and ATP levels were significantly better than 10% BSA control group (P<0.05). In conclusion, adding NA to sheep semen low temperature dilution under 4℃ storage conditions can improve sperm motility, movement characteristic, plasma membrane integrity, antioxidant capacity, sperm mitochondrial function and ATP. It can effectively improve the low temperature preservation effect of sheep sperm.

Key words: sheep, nicotinic acid, sperm motility, mitochondria, oxidative stress

中图分类号:

S826.34

韩修远, 赵亮, 王闯, 亓美玉, 姚玉昌. 烟酸通过降低氧化应激水平提高绵羊精子低温保存效果[J]. 畜牧兽医学报, 2023, 54(5): 1979-1989.

HAN Xiuyuan, ZHAO Liang, WANG Chuang, QI Meiyu, YAO Yuchang. Nicotinic Acid Enhances Low Temperature Preservation of Sheep Sperm by Reducing Oxidative Stress Levels[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 1979-1989.

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