N,N-二甲基甲酰胺(DMF)对猪精液冷冻效果的影响

Effect of N,N-Dimethylformamide on cryopreservation of boar semen

【目的】研究N,N-二甲基甲酰胺(N,N-Di methylformamide,DMF)与甘油配制的混合保护剂对猪冷冻精子的保护效果。【方法】以DMF和甘油配制成混合冷冻保护剂对猪精液进行冷冻,试验共设对照组(30mL/L甘油)、A组(10mL/L DMF+20mL/L甘油)、B组(20mL/L DMF+10mL/L甘油)、C组(30mL/L DMF)和D组(50mL/L DMF)5个处理组,以活动精子百分率、精子活率、精子直线运动速率、精子顶体完整率以及精子质膜完整率为检测指标,对猪精子冷冻效果进行评价。【结果】20mL/L DMF+10mL/L甘油混合冷冻保护剂对猪精子有较好的冷冻保护作用,与其他处理组相比,除精子直线运动速率和质膜完整率与10mL/L DMF+20mL/L甘油组无显著差异外,其他各项指标均优于其他组及甘油对照组。30mL/L DMF对猪精子的毒性强于同浓度甘油,但20mL/LDMF+10mL/L甘油混合冷冻保护剂的毒性与甘油对照组无显著差异(P>0.05)。【结论】20mL/L DMF+10mL/L甘油混合冷冻保护剂比单一使用甘油或DMF能起到更好的冷冻保护作用。

[Objective] The effect of mixed cryoprotectants composed of different proportions of glycerol and N,N-Dimethylformamide （DMF） on freezing boar sperm was studied. [Method] The mixing cryoprotectant was composed of DMF and glycerol,and experiments were executed among five treatments： the control （30 mL/L glycerol）, group A （10 mL/L DMF ＋20 mL/L glycerol）, group B （20 mL/L DMF ＋ 10 mL/L glycerol）,group C （30 mL/L DMF） and group D （50 mL/L DMF）. In the present study, the effects of mixing cryoprotectant on frozen-thawed sperm were evaluated by five sperm parameters including the percentage of total motile sperm, motility, velocity of straight-line motion, plasma membrane integrity and acrosome integrity. [Result] 20 mL/L DMF ＋ 10 mL/L glycerol could protect boar sperm against cryodamage during cryopreservation. No difference on velocity of straight-line motion and membrane integrity of frozen-thawed sperm was found between group B and group A （P〉0.05）, and the parameters of frozen-thawed sperm of the group B were higher than those of other groups （P〈0.05）;The toxicity of DMF was stronger than that of glycerol at the same level of concentration （30 mL/L）, and no difference existed between group B and the control in the present study （P〈0.05）. [Conclusion] 20 mL/L DMF ＋10 mL/L glycerol could provide better protective action for frozen boar sperm than that of DMF or glycerol alone.