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超氧化物歧化酶(SOD)蛋白对精液稀释液中马精子活力、存活、顶体状态和细胞外信号调节激酶(ERK)蛋白磷酸化的影响。

Effect of sod (superoxide dismutase) protein supplementation in semen extenders on motility, viability, acrosome status and ERK (extracellular signal-regulated kinase) protein phosphorylation of chilled stallion spermatozoa.

机构信息

Department of Veterinary Clinical Sciences, University of Naples Federico II, Naples, Italy.

出版信息

Theriogenology. 2011 Apr 15;75(7):1201-10. doi: 10.1016/j.theriogenology.2010.11.031. Epub 2011 Feb 4.

Abstract

New studies are underway to find new methods for supporting longer storage of cooled stallion semen. It is known that high concentrations of reactive oxygen species (ROS) cause sperm pathology. The metalloprotein superoxide dismutase (SOD) is responsible for H(2)O(2) and O(2) production, by dismutation of superoxide radicals. The aim of this study is to assess the quality of chilled stallion semen processed with extenders containing SOD at different concentrations as antioxidant additives. A total of 80 ejaculates collected from 5 standardbred stallions was divided into 5 aliquots treated as: native semen (control 1); native semen diluted 1:3 with Kenney semen extender (control 2); spermatozoa diluted after centrifugation in extender without (control 3) or with SOD at 25 IU/ml (experimental 1) or 50 IU/ml (experimental 2). Each sample was analyzed for motility, viability and acrosome status, immediately after semen preparation and again after storage at 5 °C for 24 h, 48 h and 7 2h. Acrosome integrity was evaluated by Chlortetracycline (CTC) and Fluorescent-labeled peanut lectin agglutinin (PNA-FITC conjugated staining). A proteomic approach of quantifying extracellular signal regulated kinase (ERK) was also evaluated as an indirect indicator of oxidative stress. In all samples sperm progressive motility and sperm acrosomal integrity showed a significant reduction between fresh and cooled spermatozoa at 24 h, 48 h and 72 h. Quality parameters of sperm were significantly higher (Progressive Motility P < 0.01; Viability P < 0.001) in aliquots supplemented with SOD. ERK phosphorylation was statistically higher (P < 0.01) in aliquots without SOD. The Authors concluded that addition of SOD to semen extenders improves the quality of chilled equine semen and reduces ERK activation.

摘要

正在进行新的研究,以寻找支持冷却保存种马精液的新方法。已知高浓度的活性氧物种 (ROS) 会导致精子病变。金属蛋白酶超氧化物歧化酶 (SOD) 通过歧化超氧自由基,负责 H(2)O(2) 和 O(2) 的产生。本研究旨在评估含有不同浓度 SOD 的稀释剂处理后的冷却种马精液的质量,作为抗氧化添加剂。从 5 匹标准种马收集的 80 份精液分为 5 份:原精液(对照 1);原精液用 Kenney 精液稀释剂稀释 1:3(对照 2);精子经离心后在无(对照 3)或 25 IU/ml(实验 1)或 50 IU/ml(实验 2)SOD 的稀释剂中处理。每个样本均在精液准备后立即进行运动性、活力和顶体状态分析,并在 5°C 储存 24 小时、48 小时和 72 小时后再次分析。顶体完整性通过氯四环素 (CTC) 和荧光标记花生凝集素 (PNA-FITC 结合染色) 进行评估。还评估了定量细胞外信号调节激酶 (ERK) 的蛋白质组学方法,作为氧化应激的间接指标。在所有样本中,新鲜和冷却精子在 24 小时、48 小时和 72 小时时,精子的运动性和精子顶体完整性均显著降低。添加 SOD 的样本的精子质量参数(精子运动性 P < 0.01;活力 P < 0.001)显著更高。无 SOD 的样本中 ERK 磷酸化统计学上更高(P < 0.01)。作者得出结论,向精液稀释剂中添加 SOD 可提高冷却马精液的质量并减少 ERK 激活。

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