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半胱氨酸和超氧化物歧化酶对解冻后鸡精子质量的影响。

The effect of cysteine and superoxide dismutase on the quality of post-thawed chicken sperm.

机构信息

Wroclaw University of Environmental and Life Sciences, Faculty of Veterinary Medicine, Department of Reproduction and Clinic of Farm Animals, pl. Grunwaldzki 49, 50-366 Wrocław, Poland.

出版信息

Cryobiology. 2013 Oct;67(2):132-6. doi: 10.1016/j.cryobiol.2013.06.002. Epub 2013 Jun 12.

Abstract

The study was conducted to determine the influence of N-acetyl-l-cysteine (NAC) and superoxide dismutase (SOD) on chicken sperm motility, plasma membrane and acrosome integrity, mitochondrial activity, lipid peroxidation (LPO) and apoptotic changes after freezing-thawing process. Semen samples from fifteen Greenlegged Partridge roosters were pooled, diluted with EK extender without antioxidants (control), or supplemented with 5 mM NAC, or 200 U/mL SOD. Samples were subjected to cryopreservation. After thawing, sperm parameters evaluated by using CASA system and flow cytometry were assessed. The extender supplemented with NAC and SOD caused the increase (P < 0.01) in sperm motility and provided the higher percentage of rapid sperm (P < 0.01) compared to control. The addition of NAC increased the progressive motility of cells (P < 0.01). In NAC and SOD groups post-thaw plasma membrane integrity was higher (P < 0.05) and less spermatozoa showed apoptotic changes (P < 0.01, P < 0.05). Post-thaw percentage of sperm with high mitochondrial activity was the greatest (P < 0.05) with NAC addition. The SOD supplementation only reduced (P < 0.05) the percentage of sperm with LPO, following the cryopreservation. These results indicate that the addition of NAC (5 mM) and SOD (200 U/mL) is beneficial for the function of frozen-thawed chicken spermatozoa. The antioxidants prevented the reduction in motility, viability and mitochondrial membrane potential, as well as protected from apoptotic changes in sperm. Lipid peroxidation in sperm plasma membrane was decreased by SOD supplementation. Therefore, these antioxidants can be recommended as an additional component of chicken freezing extender.

摘要

本研究旨在探讨 N-乙酰半胱氨酸(NAC)和超氧化物歧化酶(SOD)对鸡精子运动能力、质膜和顶体完整性、线粒体活性、脂质过氧化(LPO)和冻融后凋亡变化的影响。从 15 只绿胫鹧鸪公鸡中收集精液样本,将其与不含抗氧化剂的 EK 稀释液(对照组)或添加 5mM NAC 或 200U/mL SOD 的稀释液混合。将样品进行冷冻保存。解冻后,使用 CASA 系统和流式细胞术评估精子参数。与对照组相比,添加 NAC 和 SOD 的稀释液可提高精子活力(P<0.01),并提供更高比例的快速精子(P<0.01)。添加 NAC 可提高细胞的运动速度(P<0.01)。在 NAC 和 SOD 组中,解冻后质膜完整性更高(P<0.05),凋亡变化的精子数量更少(P<0.01,P<0.05)。添加 NAC 后,解冻后具有高线粒体活性的精子比例最大(P<0.05)。只有 SOD 补充剂可降低(P<0.05)冷冻保存后精子的 LPO 百分比。这些结果表明,添加 NAC(5mM)和 SOD(200U/mL)有利于冷冻解冻鸡精子的功能。抗氧化剂可防止精子活力、活力和线粒体膜电位降低,并防止精子发生凋亡变化。SOD 补充剂可降低精子质膜的 LPO。因此,这些抗氧化剂可作为鸡冷冻稀释液的附加成分推荐使用。

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