在公羊精子冷冻保存培养基中添加Mn、Cu、Zn纳米琥珀酸盐后精子的质量和受精能力

O. Sharan, V. Stefanyk, D. Ostapiv
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引用次数: 0

摘要

在精子冷冻保存过程中,精子的超微结构受到破坏,导致精子的生化和功能发生变化。为了保护精子免受低温的负面影响,使用了添加微量元素的低温保存培养基。最近,在乌克兰获得了微量元素的纳米形式,并开始实验研究其对动物身体的影响。研究了在公羊精子冷冻保存培养基中添加锰、锌、铜纳米琥珀酸盐对精子质量和受精率的影响。试验选用6只临床健康的2 ~ 4岁种公羊。选取6只2 ~ 4岁的特塞尔公羊进行射精评价,分为对照组和试验组。对照精子样品用乳糖-蛋黄-三柠檬酸盐-甘油培养基(LYTCGM)稀释。将微量元素纳米琥珀酸盐分别以Zn和Mn - 2.5、5.0和7.5 μg/l、Cu - 1.25、2.5和3.75 μg/l的剂量添加到公羊精子样品的培养基中。稀释后的精子用吸管包装,平衡2.5小时后冷冻。精子解冻后,测定精子活力、受损精子百分比、精子存活率、琥珀酸脱氢酶(SDH)和细胞色素氧化酶(CO)活性。在LYTCGM中加入锰、锌和铜纳米柠檬酸盐,形成了剂量依赖性效应。在LYTCGM中添加剂量为5.0 μg/l的纳米琥珀酸锰和锌,可能会增加公羊精子解冻后的活力,并降低形态障碍精子的比例。增加纳米琥珀酸铜的添加量显著降低了解冻公羊精液中的精子活力,同时增加了退化精子的百分比。以5.0 μg/l LYTCGM剂量添加纳米琥珀酸锰和锌后,公羊精子的存活率可能有所提高,而纳米琥珀酸铜的添加剂量越高,生殖细胞的存活时间越短。在LYTCGM中添加5.0 μg/l的纳米琥珀酸锰和锌可能会增加解冻后精子中SDH和CO的活性,而添加越来越多的纳米琥珀酸铜则会显著降低这些酶的活性。低剂量(2.5 μg/l)和高剂量(7.5 μg/l)添加纳米琥珀酸锰和锌对精子运动、形态障碍和存活以及SDH和CO活性均无显著影响。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
The quality and fertilizing ability of sperm after the addition of nanosuccinates of Mn, Cu, Zn to the medium for cryopreservation of ram sperm
In the process of cryopreservation of sperm, there are violations of the ultrastructure of spermatozoa, which causes biochemical and functional changes in them. To protect spermatozoa from the negative effects of low temperatures, cryopreservation media are used, to which trace elements are added. Recently, nanoforms of trace elements have been obtained in Ukraine and experiments have been started to study their effect on the animal body. The aim of the work was to find out the effect of adding nanosuccinate of Mn, Zn and Cu to the medium for cryopreservation of ram sperm on the quality and fertilizing ability of sperm. The experiment was conducted on six clinically healthy breeder rams, aged 2–4 years. After evaluating the ejaculate from six rams aged 2–4 years, the Texel breed was divided into control and experimental groups. Control sperm samples were diluted with lactose-yolk-tris-citrate-glycerin medium (LYTCGM). Nanosuccinates of microelements were added to the medium in test samples of ram sperm in the following doses: Zn and Mn – 2.5, 5.0 and 7.5 μg/l, Cu – 1.25, 2.5 and 3.75 μg/l. Diluted sperm was packaged in straws, equilibrated for 2.5 hours and frozen. After thawing of sperm, motility, percentage of damaged spermatozoa, their survival, activity of succinate dehydrogenase (SDH) and cytochrome oxidase (CO) in sperm were determined. A dose-dependent effect of Mn, Zn, and Cu nanocitrates upon their addition to LYTCGM was established. The addition of Mn and Zn nanosuccinate at a dose of 5.0 μg/l to LYTCGM probably increases the motility of ram spermatozoa after thawing, and also reduces the percentage of spermatozoa with morphological disorders. Addition of Cu nanosuccinate in increasing doses significantly reduces spermatozoa motility in thawed ram semen, simultaneously increasing the percentage of degenerate spermatozoa. After the addition of Mn and Zn nanosuccinate at a dose of 5.0 μg/l LYTCGM, the survival rate of ram spermatozoa is probably increased, and the addition of Cu nanosuccinate in increasing doses significantly reduces the survival time of germ cells. Addition of Mn and Zn nanosuccinate at a dose of 5.0 μg/l to LYTCGM probably increases the activity of SDH and CO in sperm after thawing, and the addition of Cu nanosuccinate in increasing doses significantly reduces the activity of these enzymes. The addition of Mn and Zn nanosuccinate in lower (2.5 μg/l) and higher (7.5 μg/l) doses did not significantly affect the motility, morphological disorders and survival of spermatozoa, as well as the activity of SDH and CO in them.
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