Zhu Zhendong, Ren Zhanjun, Fan Xiaoteng, Pan Yang, Lv Shan, Pan Chuanying, Lei Anmin, Zeng Wenxian
College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, China.
College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
PLoS One. 2017 Jul 10;12(7):e0181110. doi: 10.1371/journal.pone.0181110. eCollection 2017.
The process of cryopreservation results in over-production of reactive oxygen species, which is extremely detrimental to spermatozoa. The aim of this study was to investigate whether addition of cysteine to freezing extender would facilitate the cryosurvival of rabbit spermatozoa, and if so, how cysteine protects spermatozoa from cryodamages. Freshly ejaculated semen was diluted with Tris-citrate-glucose extender supplemented with different concentrations of cysteine. The motility, intact acrosomes, membrane integrity, mitochondrial potentials, 8-hydroxyguanosine level and sperm-zona pellucida binding capacity were examined. Furthermore, glutathione peroxidase (GPx) activity, glutathione content (GSH), and level of reactive oxygen species (ROS) and hydrogen peroxide of spermatozoa were analyzed. The values of motility, intact acrosomes, membrane integrity, mitochondrial potentials and sperm-zona pellucida binding capacity of the frozen-thawed spermatozoa in the treatment of cysteine were significantly higher than those of the control. Addition of cysteine to extenders improved the GPx activity and GSH content of spermatozoa, while lowered the ROS, DNA oxidative alterations and lipid peroxidation level, which makes spermatozoa avoid ROS to attack DNA, the plasma membrane and mitochondria. In conclusion, cysteine protects spermatozoa against ROS-induced damages during cryopreservation and post-thaw incubation. Addition of cysteine is recommended to facilitate the improvement of semen preservation for the rabbit breeding industry.
冷冻保存过程会导致活性氧的过度产生,这对精子极其有害。本研究的目的是探讨在冷冻稀释液中添加半胱氨酸是否会促进兔精子的冷冻存活,如果是,半胱氨酸如何保护精子免受冷冻损伤。用添加不同浓度半胱氨酸的Tris-柠檬酸-葡萄糖稀释液稀释新鲜射出的精液。检测精子活力、顶体完整率、膜完整性、线粒体膜电位、8-羟基鸟苷水平和精子-透明带结合能力。此外,还分析了精子的谷胱甘肽过氧化物酶(GPx)活性、谷胱甘肽含量(GSH)、活性氧(ROS)水平和过氧化氢水平。半胱氨酸处理组冻融后精子的活力、顶体完整率、膜完整性、线粒体膜电位和精子-透明带结合能力均显著高于对照组。向稀释液中添加半胱氨酸可提高精子的GPx活性和GSH含量,同时降低ROS水平、DNA氧化改变和脂质过氧化水平,从而使精子避免ROS对DNA、质膜和线粒体的攻击。总之,半胱氨酸可保护精子在冷冻保存和解冻后孵育过程中免受ROS诱导的损伤。建议添加半胱氨酸以促进兔养殖业精液保存的改善。