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海藻糖和己酮可可碱对山羊精液解冻过程中冷胁迫的保护作用。

Protective Effects of Trehalose and Pentoxifylline on Goat Sperm Exposed to Chilling-Freezing Process.

机构信息

Laboratory of Reproduction Biology, Department of Animal Science, University of Kurdistan, Sanandaj, Iran.

出版信息

Biopreserv Biobank. 2022 Dec;20(6):540-550. doi: 10.1089/bio.2021.0081. Epub 2022 Jan 7.

DOI:10.1089/bio.2021.0081
PMID:35005998
Abstract

Freezing of sperm is known as an important part of assisted reproduction. However, many studies have illustrated that cryopreservation negatively affects the quality and fertility rate of sperm. This study aimed to evaluate the effects of trehalose and pentoxifylline (PTX) in diluents on cooled and frozen-thawed Markhoz goat sperm. Preassessed samples were pooled and diluted with a basic diluent using trehalose and PTX. The cooled sperm showed significant improvement. The motion characteristics of cryopreserved sperm were evaluated based on computer-assisted system analysis. In this study, we investigated the viability, membrane integrity, malondialdehyde concentration, total abnormality, acrosome integrity, and seminal hyaluronidase enzyme. Also, the hypo-osmotic swelling test, mitochondrial activity, apoptotic features, caspase activity, chromatin dispersion test, active mitochondria, and reactive oxygen species (ROS) activity were assessed as complementary parameters. The data illustrate that the total motility, progressive motility, average path velocity (VAP), straight-line velocity (VSL), curvilinear velocity (VCL), and the ratio of sperm chromatin dispersion, viable sperm were improved significantly ( < 0.05) using 3 mM PTX alone or 3 mM PTX plus 50 mM trehalose, while other characteristics indicate significant enhancement by 3 and 6 mM PTX and 50 and 70 mM trehalose alone or in combination, except amplitude of lateral head displacement (ALH), beat/cross frequency (BCF), and intracellular ROS-(O), which demonstrate no significant difference among treatments. In conclusion, this study indicates that addition of 3 and 6 mM PTX alone or with 50 and 70 mM trehalose seems to reduce the damage caused by cooling and cryopreservation processes.

摘要

精子冷冻保存作为辅助生殖的重要组成部分已经广为人知。然而,许多研究表明,冷冻保存会对精子的质量和生育能力产生负面影响。本研究旨在评估海藻糖和己酮可可碱(PTX)在稀释剂中对冷却和冻融的 Markhoz 山羊精子的影响。预先评估的样本进行混合,并用基本稀释剂与海藻糖和 PTX 混合。冷却后的精子显示出显著的改善。通过计算机辅助系统分析评估冷冻保存精子的运动特征。在这项研究中,我们调查了活力、膜完整性、丙二醛浓度、总畸形率、顶体完整性和精液透明质酸酶。还评估了低渗肿胀试验、线粒体活性、凋亡特征、半胱天冬酶活性、染色质弥散试验、活性线粒体和活性氧(ROS)活性作为补充参数。数据表明,单独使用 3mM PTX 或 3mM PTX 加 50mM 海藻糖可显著提高总活力、前向活力、平均路径速度(VAP)、直线速度(VSL)、曲线速度(VCL)和精子染色质弥散的比例,有活力的精子显著提高(<0.05),而其他特征表明,单独使用 3 和 6mM PTX 以及 50 和 70mM 海藻糖,或联合使用时,均显著增强,除侧向头部位移振幅(ALH)、拍动/交叉频率(BCF)和细胞内 ROS-(O)外,这些参数在处理组之间没有显著差异。总之,本研究表明,单独添加 3 和 6mM PTX 或与 50 和 70mM 海藻糖混合似乎可以减少冷却和冷冻保存过程造成的损伤。

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