海藻糖对猪精液冷冻保存效果的影响

Effects of Trehalose on Cryopreservation of Boar Semen

在传统的Tris-柠檬酸-葡萄糖稀释液基础上,分别添加25%、50%、75%、100%的海藻糖,研究不同浓度海藻糖对猪精液冷冻后精子质量的影响。结果表明,海藻糖相对于对照TCG稀释液能够显著改善和提高猪精液的冷冻效果,其最佳添加浓度为25%,冷冻-解冻后猪精子活力、活率、线粒体活性、质膜完整性以及顶体完整率均显著提高(P<0.05),分别达到41.38%、46.34%、44.56%、43.51%和64.09%。海藻糖可以明显抑制精子获能,获能处理前精子获能率仅为3.68%,而获能处理后达到41.82%,有利于促进精子获能。精液稀释液中甘油的适宜添加浓度为2%,海藻糖只有与甘油共同作用,才能在冷冻-解冻过程更加有效地保护精子。猪精子活力、活率、线粒体活性、质膜完整率、顶体完整率等之间存在极显著的正相关关系(P<0.01),而与获能处理前精子的获能率存在显著的负相关关系(P<0.05)。

In order to improve boar sperm quality in frozen-thawed process, trehalose was added into the basic extenders-Tris-Citric acid-Glucose. The effects of trehalose on quality of cryopreservation of boar sperm were investigated. Moreover, in order to determine the optimum concentration of trehalose, we designed four different concentrations.25% ,50% ,75% and 100%（v/ v）, respectively. The results showed that the effects of the basic extenders supplemented with trehalose on cryoprotection of spermatozoa were better than that of traditional Tris-Citric acidGlucose （TCG） without supplement. We found out that TCG extender with 25% trehalose concentration significantly enhanced motility, viability, mitochondrial activity, membrane integrity and acrosome integrity of boar sperm after freezing and thawing （P〈 0.05）, and they were 41.38%,46.34%,44.56%,43.51% and 64.09%, respectively. Extender containing 25% trehalose could enable some improvement of cryopreservation of boar semen over the currently used TCG medium. Trehalose could decrease distinctly the percentage of spermatozoa capacitated caused by freezing and thawing（3.68%）. Capacitation potential was improved significantly with the addition of trehalose in the diluents, especially at 25% concentration（41.82%）. The optimal concentration of glycerol in extender was found to be 2% （v/v）. Trehalose could improve spermatozoa quality after freeze-thaw process when there was glycerol in the extender. The results of present study indicated that there was a significant positive correlation among sperm motility, viability, mitochondrial activity, membrane integrity and acrosome integrity（P〈0.01）. However, the percentage of spermatozoa capacitated before capacitation treatment was highly negative correlated with sperm quality index in this study（P〈0. 05）.