Basioura A, Tsakmakidis I A, Martinez E A, Roca J, Li J, Molina M F, Theodoridis A, Boscos C M, Parrilla I
Unit of Biotechnology of Reproduction, Farm Animals Clinic, School of Veterinary Medicine, Faculty of Health Sciences, Aristotle University of Thessaloniki, 54627, Thessaloniki, Greece.
Unit of Biotechnology of Reproduction, Farm Animals Clinic, School of Veterinary Medicine, Faculty of Health Sciences, Aristotle University of Thessaloniki, 54627, Thessaloniki, Greece.
Anim Reprod Sci. 2020 Jul;218:106478. doi: 10.1016/j.anireprosci.2020.106478. Epub 2020 May 1.
The aim of the study was to determine whether the presence of astaxanthin (ASX) protects boar spermatozoa against damage related to cryopreservation. Pooled ejaculates extended in Beltsville Thawing Solution (BTS) were used. Three experiments were conducted: 1) sperm samples were pre-incubated overnight (17 °C) with ASX (0, 0.5, 5, 15 μM) prior to freezing and then frozen using cooling and thawing extenders supplemented with ASX (0, 0.5, 5, 15 μM); 2) sperm samples were treated with ASX (0, 0.5, 5, 15 μM) only during overnight pre-incubation (17 °C) prior to cryopreservation; and 3) a thawing extender was supplemented with ASX (0, 0.5, 5, 15 μM). The groups were as follows: control (C; no treatment), ASX 1 (0.5 μM), ASX 2 (5 μM) and ASX 3 (15 μM). Total (TM) and progressive (PM) motility was analyzed using CASA, while sperm viability, reactive oxygen species generation, lipid peroxidation and apoptoticlike changes were analyzed using flow cytometry. Sperm variables were evaluated prior to freezing as well as 30 and 150 min after thawing. In Experiment 1, the values of TM and sperm viability post-thaw were less in the ASX 3 than C group. In Experiment 2, there was no effect of ASX on any of the sperm variables evaluated, while in Experiment 3, apoptotic-like changes were less in the ASX 1 than C group. In conclusion, there was a subtle beneficial effect on cryopreserved boar spermatozoa after addition of ASX to thawing media.
本研究的目的是确定虾青素（ASX）的存在是否能保护公猪精子免受冷冻保存相关的损伤。使用在贝尔茨维尔解冻液（BTS）中稀释的混合射精样本。进行了三项实验：1）精子样本在冷冻前于17°C下与ASX（0、0.5、5、15μM）预孵育过夜，然后使用补充有ASX（0、0.5、5、15μM）的冷冻和解冻稀释液进行冷冻；2）精子样本仅在冷冻保存前的过夜预孵育（17°C）期间用ASX（0、0.5、5、15μM）处理；3）在解冻稀释液中补充ASX（0、0.5、5、15μM）。分组如下：对照组（C；未处理）、ASX 1（0.5μM）、ASX 2（5μM）和ASX 3（15μM）。使用计算机辅助精子分析（CASA）分析总活力（TM）和渐进活力（PM），同时使用流式细胞术分析精子活力、活性氧生成、脂质过氧化和凋亡样变化。在冷冻前以及解冻后30和150分钟评估精子变量。在实验1中，解冻后ASX 3组的TM值和精子活力低于C组。在实验2中，ASX对所评估的任何精子变量均无影响，而在实验3中，ASX 1组的凋亡样变化低于C组。总之，在解冻培养基中添加ASX后，对冷冻保存的公猪精子有微妙的有益作用。
