Catalán Jaime, Yánez-Ortiz Iván, Torres-Garrido Marc, Ribas-Maynou Jordi, Llavanera Marc, Barranco Isabel, Yeste Marc, Miró Jordi
Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, ES-17003 Girona, Spain.
Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, ES-17003 Girona, Spain.
Antioxidants (Basel). 2024 Mar 6;13(3):322. doi: 10.3390/antiox13030322.
Cryopreservation is a stressful process for sperm, as it is associated with an increased production of reactive oxygen species (ROS). Elevated ROS levels, which create an imbalance with antioxidant capacity, may result in membrane lipid peroxidation (LPO), protein damage and DNA fragmentation. This study aimed to determine whether the membrane LPO and DNA fragmentation of frozen-thawed horse sperm relies upon antioxidant activity, including enzymes (superoxide dismutase (SOD), glutathione peroxidase (GPX), catalase (CAT) and paraoxonase type 1 (PON1)); non-enzymatic antioxidant capacity (Trolox-equivalent antioxidant capacity (TEAC), plasma ferric reducing antioxidant capacity (FRAP) and cupric reducing antioxidant capacity (CUPRAC)); and the oxidative stress index (OSI) of their seminal plasma (SP). Based on total motility and plasma membrane integrity (SYBR14/PI) after thawing, ejaculates were hierarchically ( < 0.001) clustered into two groups of good- (GFEs) and poor-(PFEs) freezability ejaculates. LPO and DNA fragmentation (global DNA breaks) were higher ( < 0.05) in the PFE group than in the GFE group, with LPO and DNA fragmentation (global DNA breaks) after thawing showing a positive relationship ( < 0.05) with SP OSI levels and ROS production. In addition, sperm motility and membrane integrity after thawing were negatively ( < 0.05) correlated with the activity levels of SP antioxidants (PON1 and TEAC). The present results indicate that LPO and DNA fragmentation in frozen-thawed horse sperm vary between ejaculates. These differences could result from variations in the activity of antioxidants (PON1 and TEAC) and the balance between the oxidant and antioxidant components present in the SP.
冷冻保存对精子来说是一个应激过程，因为它与活性氧（ROS）的产生增加有关。升高的ROS水平会导致抗氧化能力失衡，可能会导致膜脂质过氧化（LPO）、蛋白质损伤和DNA片段化。本研究旨在确定冻融马精子的膜LPO和DNA片段化是否依赖于抗氧化活性，包括酶（超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GPX）、过氧化氢酶（CAT）和1型对氧磷酶（PON1））；非酶抗氧化能力（Trolox等效抗氧化能力（TEAC）、血浆铁还原抗氧化能力（FRAP）和铜还原抗氧化能力（CUPRAC））；以及它们精浆（SP）的氧化应激指数（OSI）。根据解冻后的总活力和质膜完整性（SYBR14/PI），射精样本被分层（<0.001）聚为两组，即冷冻保存性好的射精样本（GFEs）和冷冻保存性差的射精样本（PFEs）。PFE组的LPO和DNA片段化（整体DNA断裂）高于（<0.05）GFE组，解冻后的LPO和DNA片段化（整体DNA断裂）与SP的OSI水平和ROS产生呈正相关（<0.05）。此外，解冻后的精子活力和膜完整性与SP抗氧化剂（PON1和TEAC）的活性水平呈负相关（<0.05）。目前的结果表明，冻融马精子中的LPO和DNA片段化在不同射精样本间存在差异。这些差异可能是由于抗氧化剂（PON1和TEAC）活性的变化以及SP中氧化剂和抗氧化剂成分之间的平衡所致。
