线粒体效应,DNA损伤和抗氧化酶活性在冷冻保存的人类精子样本:一项试点研究

P. Pinto-Pinho, R. Arantes-Rodrigues, I. Gaivão, Francisco Peixoto, Z. Gomes, M. Brito, O. Moutinho, B. Colaço, Rosário Pinto-Leite
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引用次数: 3

摘要

本研究旨在评估蒸汽冷冻方案对抗氧化酶活性(超氧化物歧化酶(SOD)和谷胱甘肽还原酶(GR))、线粒体活性精子、DNA损伤和精子参数的影响。用(方法A)和不(方法B)分别用商业精子冷冻培养基(SFM)和用精子制备培养基(SPM)进行固有去除,将26份不育男性的精液样本冷冻保存在液氮(LN)蒸气中,然后注入液氮(LN)。除SOD和GR活性仅在冻结后进行评估外,其余参数均在冻结前后进行评估。虽然A方法的效果好于B方法,但线粒体活跃、活力、活力和形态正常的精子比例显著降低。两种方法的DNA损伤(通过彗星法测定)都相似地增加,但当采用方法A时,DNA片段化的精子百分比(通过TUNEL法测定)保持与新鲜值相似。方法a的GR活性较高,SOD活性较低。添加SFM与SPM洗精似乎是保持大多数分析精子参数和活性线粒体质量的必要条件。根据相关系数,冷冻造成的有害影响取决于新鲜精液的质量。有趣的是,两种方法的解冻样本都有相似的DNA损伤。这些结果强调需要找到更有效的方案,特别是对于低质量精液样本的冷冻。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Mitochondrial Effects, DNA Damage, and Antioxidant Enzyme Activity in Cryopreserved Human Sperm Samples: A Pilot Study
This study aimed to evaluate the impact of a vapor freezing protocol on antioxidant enzyme activity (superoxide dismutase (SOD) and glutathione reductase (GR)), sperm with active mitochondria, DNA damage, and spermatic parameters. Twenty-six semen samples from men undergoing infertility investigation were cryopreserved in liquid nitrogen (LN) vapors and plunged into LN, with (method A) and without (method B) a commercial sperm freezing medium (SFM) and inherent removal with a sperm preparation medium (SPM). Most parameters were assessed before and after freezing, except for SOD and GR activity, which were only assessed after freezing. Although method A promoted better results than method B, the percentage of spermatozoa with active mitochondria, motility, vitality, and normal morphology decreased significantly. DNA damage (determined by comet assay) increased similarly with both methods, but the percentage of spermatozoa with fragmented DNA (by TUNEL assay) remained similar to fresh values when method A was applied. GR activity was higher and SOD activity lower with method A. The addition of SFM coupled with the sperm wash with SPM seems essential to preserve the quality of most of the analyzed spermatic parameters and active mitochondria. The detrimental effects promoted by freezing were shown to depend on the quality of the fresh semen, according to correlation coefficients. Interestingly, thawed samples of both methods shared similar DNA damage. These results highlight the need to find more effective protocols, especially for the freezing of low-quality semen samples.
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