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冷冻保护剂对俄罗斯鲟(Acipenser ruthenus)精子质量、DNA完整性、抗氧化反应及氧化应激抗性的影响

The influence of cryoprotectants on sturgeon (Acipenser ruthenus) sperm quality, DNA integrity, antioxidant responses, and resistance to oxidative stress.

作者信息

Shaliutina-Kolešová Anna, Cosson Jacky, Lebeda Ievgen, Gazo Ievgenia, Shaliutina Olena, Dzyuba Boris, Linhart Otomar

机构信息

University of South Bohemia in České Budějovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Research Institute of Fish Culture and Hydrobiology, Zátiší 728/II, 389 25 Vodňany, Czech Republic.

University of South Bohemia in České Budějovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Research Institute of Fish Culture and Hydrobiology, Zátiší 728/II, 389 25 Vodňany, Czech Republic.

出版信息

Anim Reprod Sci. 2015 Aug;159:66-76. doi: 10.1016/j.anireprosci.2015.05.014. Epub 2015 May 22.

Abstract

This study examined the effect of cryoprotectants on DNA integrity, antioxidant defense, and resistance to oxidative stress in cryopreserved sterlet Acipenser ruthenus sperm. The freeze-thaw process significantly influenced sperm motility, with significant differences among cryoprotectants. In vitro exposure of cryopreserved sperm to the xanthine-xanthine oxidase (X-XO) system as a model reactive oxygen species inducer resulted in a lesser motility rate and velocity compared to the control, and there was a decrease in these variables in a time- and dose-dependent manner. The greatest X (0.6mM)-XO (0.05U/mL) concentration and incubation period (30min) was associated with 62% DNA fragmentation in sperm cryopreserved with 10% ethylene glycol (EG). The maximum lipid peroxidation (LPO) and carbonyl derivatives of proteins (CP) was also observed in sperm cryopreserved with 10% EG and exposed to the X-XO system at a concentration of 0.6mM X-0.05U/mL XO. The frozen/thawed sperm containing 10% EG and that with 10% dimethyl sulfoxide (DMSO) had a significant enhancement of superoxide dismutase (SOD) and glutathione reductase (GR) activity. The current study confirms that EG is not effective for cryopreservation, and sterlet sperm were highly sensitive to free radicals after cryopreservation with EG.

摘要

本研究考察了冷冻保护剂对冷冻保存的俄罗斯鲟精子DNA完整性、抗氧化防御及氧化应激抗性的影响。冻融过程显著影响精子活力,不同冷冻保护剂之间存在显著差异。将冷冻保存的精子体外暴露于作为活性氧诱导模型的黄嘌呤-黄嘌呤氧化酶(X-XO)系统中,与对照组相比,精子活力率和速度降低,且这些变量随时间和剂量呈依赖性降低。在用10%乙二醇(EG)冷冻保存的精子中,最大的X(0.6mM)-XO(0.05U/mL)浓度和孵育时间(30分钟)与62%的DNA片段化相关。在用10%EG冷冻保存并暴露于浓度为0.6mM X-0.05U/mL XO的X-XO系统的精子中,还观察到最大的脂质过氧化(LPO)和蛋白质羰基衍生物(CP)。含有10%EG和10%二甲基亚砜(DMSO)的冻融精子超氧化物歧化酶(SOD)和谷胱甘肽还原酶(GR)活性显著增强。本研究证实,EG对冷冻保存无效,并且用EG冷冻保存后,俄罗斯鲟精子对自由基高度敏感。

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