Department of Theriogenology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.
Department of Internal Medicine and Clinical Pathology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.
Reprod Domest Anim. 2021 Jul;56(7):1004-1014. doi: 10.1111/rda.13944. Epub 2021 May 19.
Supplementing the extender with antioxidants with low molecular weight can enhance the quality of the post-thaw sperm during the freezing process. This study was aimed at determining the impacts of 3,4-dihydroxyphenyl glycol (DHPG) on the spermatozoa of the canine undergoing freeze-thawing process. In this study, 24 ejaculates were obtained from three mixed-breed dogs and were diluted in a Tris-based extender. The diluted semen was divided into aliquots for supplementation of 10, 30, 50 and 70 µg/ml of DHPG, control (without antioxidant) and control sham (DMSO). After being extended, the semen was equilibrated at a temperature of 4°C and then transferred to the straws and kept 4 cm above the liquid nitrogen for 20 min and was finally immersed in the liquid nitrogen. They were cryopreserved for seven days; then, sperm parameters including sperm motility evaluation, motility characteristics, viability, DNA and plasma membrane integrity, total antioxidant capacity (TAC), reduced glutathione content (GSH), antioxidant enzymes (superoxide dismutase [SOD], catalase [CAT] and glutathione peroxidase [GPx]) activity malondialdehyde (MDA) levels were evaluated. This study showed that spermatozoa cryopreservation with 50, 30 and 70 µg/ml of DHPG concentrations had better progressive motility, Curvilinear Velocity, Linearity, viability, intact plasma membrane and the levels of TAC, GPx and GSH were higher than the control group. The 50, 30 and 70 µg/ml of DHPG concentrations led to the significant decrease of DNA damage compared to the control group. Total motility, average path velocity, straight-line velocity and CAT activity were significantly improved in 30 and 50 µg/ml of DHPG concentrations, compared to the control group. Also, the 50 and 30 µg/ml of DHPG concentrations, decreased MDA levels compared to the other groups, significantly. In conclusion, our study showed that the addition 50 µg/ml of DHPG to the canine semen extender improved the semen characteristics and oxidative markers in the cryopreservation process.
在冷冻过程中，向延长液中添加低分子量的抗氧化剂可以提高解冻后精子的质量。本研究旨在确定 3,4-二羟基苯乙二醇（DHPG）对犬精子在冻融过程中的影响。本研究从三只杂种犬中获得 24 份精液，并在基于 Tris 的延长液中稀释。将稀释的精液分成等分试样，分别添加 10、30、50 和 70μg/ml 的 DHPG、对照（无抗氧化剂）和对照假处理（DMSO）。延长后，精液在 4°C 下平衡，然后转移到 straws 中，保持在液氮上方 4cm 处 20min，最后浸入液氮中。它们被冷冻保存七天；然后，评估精子参数，包括精子运动评估、运动特征、活力、DNA 和质膜完整性、总抗氧化能力（TAC）、还原型谷胱甘肽含量（GSH）、抗氧化酶（超氧化物歧化酶[SOD]、过氧化氢酶[CAT]和谷胱甘肽过氧化物酶[GPx]）活性和丙二醛（MDA）水平。本研究表明，精子冷冻保存浓度为 50、30 和 70μg/ml 的 DHPG 具有更好的前向运动、曲线速度、直线速度、活力、完整质膜以及 TAC、GPx 和 GSH 的水平高于对照组。与对照组相比，DHPG 浓度为 50、30 和 70μg/ml 导致 DNA 损伤显著降低。与对照组相比，DHPG 浓度为 30 和 50μg/ml 时，总运动、平均路径速度、直线速度和 CAT 活性显著提高。此外，与其他组相比，DHPG 浓度为 50 和 30μg/ml 时，MDA 水平显著降低。总之，我们的研究表明，在犬精液延长液中添加 50μg/ml 的 DHPG 可以提高冷冻保存过程中的精液特性和氧化标记物。
