Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Department of Infertility, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran.
Biopreserv Biobank. 2022 Aug;20(4):367-373. doi: 10.1089/bio.2021.0112.
Cryopreservation of spermatozoa is a general procedure to preserve viable sperm for an indefinite period. Despite the efficiency of sperm cryopreservation, excessive reactive oxygen species (ROS) production during cryopreservation can induce structural and functional changes in spermatozoa. Also, cryopreservation has been shown to decrease the spermatozoa's antioxidant activity inducing them to be more sensitive to damage caused by ROS. Experimental evidence suggests that astaxanthin (AXT) has essential activities such as antioxidant, antibacterial, and antithrombotic properties. Therefore, this study aimed to evaluate the effect of AXT on the sperm quality of healthy men during freezing-thawing. In the first phase, 10 semen samples with different concentrations of AXT (0.0, 0.5, 1, and 2 μM) were cryopreserved to achieve an optimal dose of AXT. Then, motility, viability, and phosphatidylserine (PS) externalization were evaluated. In the second phase, 25 samples were collected and divided into 3 groups: fresh group, control group (untreated frozen-thawed samples), and AXT group (treated frozen-thawed with AXT). Then, samples were cryopreserved in freezing media supplemented with or without the optimal concentration of AXT (1 μM). After thawing, the levels of sperm parameters, including motility (using a computer-assisted sperm analyzer), viability (eosin-nigrosin), early apoptotic change (annexin V/propidium iodide), ROS (flow cytometry), and lipid peroxidation (LPO) (using enzyme-linked immunosorbent assay), were evaluated. Our results showed that the addition of 1 μM AXT to sperm freezing media improved all parameters of sperm motility and viability ( ≤ 0.05). Furthermore, it could reduce the levels of ROS parameters (intracellular hydrogen peroxide and superoxide) compared with the control group ( ≤ 0.05). Also, AXT significantly decreased the level of PS externalization ( ≤ 0.05) and LPO ( ≤ 0.05) after the freezing-thawing process. In conclusion, our findings demonstrated that human semen treatment with 1 μM AXT before the freezing-thawing process has protective effects against oxidative stress and could diminish the destructive effects of this process on sperm quality.
精子的冷冻保存是一种保存有活力的精子的一般程序，可以无限期保存。尽管精子冷冻保存的效率很高，但在冷冻保存过程中过量的活性氧（ROS）的产生会导致精子结构和功能的变化。此外，冷冻保存已被证明会降低精子的抗氧化活性，使它们更容易受到 ROS 造成的损伤。实验证据表明，虾青素（AXT）具有抗氧化、抗菌和抗血栓形成等基本活性。因此，本研究旨在评估 AXT 对健康男性精子在冷冻-解冻过程中的质量的影响。在第一阶段，将 10 份精液样本用不同浓度的 AXT（0.0、0.5、1 和 2 μM）冷冻保存，以达到 AXT 的最佳剂量。然后，评估精子的活力、活力和磷脂酰丝氨酸（PS）外翻。在第二阶段，收集 25 个样本并分为 3 组：新鲜组、对照组（未经处理的冷冻-解冻样本）和 AXT 组（用 AXT 处理冷冻-解冻样本）。然后，将样本在添加或不添加最佳浓度 AXT（1 μM）的冷冻培养基中冷冻保存。解冻后，使用计算机辅助精子分析仪评估精子参数的水平，包括活力、活力（曙红-黑）、早期凋亡变化（ Annexin V/碘化丙啶）、ROS（流式细胞术）和脂质过氧化（LPO）（酶联免疫吸附试验）。我们的结果表明，在精子冷冻培养基中添加 1 μM AXT 可以改善精子活力和活力的所有参数（≤0.05）。此外，与对照组相比，它可以降低 ROS 参数（细胞内过氧化氢和超氧化物）的水平（≤0.05）。此外，AXT 还可以显著降低冷冻-解冻过程后的 PS 外翻（≤0.05）和 LPO（≤0.05）水平。总之，我们的研究结果表明，在冷冻-解冻过程之前，用 1 μM AXT 处理人类精液具有抗氧化应激的保护作用，并可以减少这个过程对精子质量的破坏性影响。
