Center of Biotechnology on Reproduction (BIOREN-CEBIOR), Faculty of Medicine, Universidad de La Frontera, Temuco, Chile.
Center of Biotechnology on Reproduction (BIOREN-CEBIOR), Faculty of Medicine, Universidad de La Frontera, Temuco, Chile; Department of Preclinical Sciences, Faculty of Medicine, Universidad de La Frontera, Temuco, Chile.
Cryobiology. 2019 Aug;89:76-81. doi: 10.1016/j.cryobiol.2019.05.003. Epub 2019 May 6.
The semen cryopreservation processes are associated with state of oxidative stress induced by high levels of reactive oxygen species (ROS), causing damage to functional spermatozoa. Whereby, antioxidants have been utilized to scavenge or neutralize the elevated levels of ROS. The aim of at the present study was to evaluate the effect of adding BHT to the freezing extenders on post-thaw characteristics of domestic cat spermatozoa. Semen samples were frozen in Tris-fructose-citric acid-based extender, supplemented with different concentrations of BHT (0.5 mM, 1.0 mM and 2.0 mM) and a control sample without antioxidant. After thawing, sperm samples were assessed for motility by computer-assisted sperm analysis and viability, acrosome integrity, superoxide anion production and membrane lipid peroxidation status by flow cytometry. In the study, the parameters of sperm motility and acrosome integrity were significantly higher in 2.0 mM BHT compared to sperm frozen in the extender with other concentrations and control (P < 0.05), in addition, this concentration reduced significantly the superoxide anion production and lipid peroxidation of the sperm. The results demonstrated that the supplementation of BHT to the freezing extender could protect the function and cellular structure of domestic cat sperm from cryoinjuries.
精液冷冻保存过程与高水平活性氧(ROS)引起的氧化应激状态有关,导致功能性精子受损。因此,抗氧化剂已被用于清除或中和升高的 ROS 水平。本研究的目的是评估在冷冻液中添加 BHT 对家猫精子解冻后特性的影响。精液样本在基于 Tris-果糖-柠檬酸的冷冻液中冷冻,添加不同浓度的 BHT(0.5mM、1.0mM 和 2.0mM)和无抗氧化剂的对照样本。解冻后,通过计算机辅助精子分析和流式细胞术评估精子活力、顶体完整性、超氧阴离子产生和膜脂质过氧化状态。在研究中,与其他浓度和对照组相比,2.0mM BHT 处理的精子在精子运动性和顶体完整性方面的参数显著更高(P<0.05),此外,该浓度还显著降低了精子的超氧阴离子产生和脂质过氧化。结果表明,在冷冻液中添加 BHT 可以保护家猫精子的功能和细胞结构免受冷冻损伤。