不同卵黄成分、大豆卵磷脂及β-环糊精胆固醇对马精子冷冻效果的影响

Effect of Different Yolk Composition,Soybean Lecithin and Cholesterol-loaded Cyclodextrin on Equine Sperm Cryopreservation

本研究旨在比较添加不同浓度的卵黄成分(全卵黄(EY)、卵黄浆蛋白(EYP)、低密度脂蛋白(LDL)及水溶性卵黄蛋白(livetin))、大豆卵磷脂(SL)及β-环糊精胆固醇(CLC)对马精子冷冻效果的影响。选择4匹成年英纯血种公马进行采精,试验1:以INRA82+3.5%的混合抗冻剂作为基础冷冻液,添加0、2%、4%和8%浓度的EY、LDL、EYP或水溶性卵黄蛋白;试验2:以4%全卵黄组为对照,添加1%、2%和4%SL;试验3:在INRA82+3.5%的混合抗冻剂+4%EY基础液中添加0、0.75、1.5和3mg/mL浓度的CLC。结果显示,EYP、LDL及水溶性卵黄蛋白3种卵黄组分均对马精子冷冻具有显著保护作用(P<0.05),但不同卵黄组分之间及与EY组相比保护效果无显著差异(P>0.05);添加低浓度(1%)的SL精子冻融后活力与对照组(4%EY组)相比无显著差异(P>0.05),但添加高浓度SL组(2%和4%)冻融后精子TM、PM及RAP值均显著低于对照组(P<0.05);添加不同浓度的CLC解冻后精子活力与未添加组无显著差异(P>0.05)。研究结果表明,卵黄中EYP、LDL和水溶性卵黄蛋白均对精子保护有效,保护效果与EY无显著差异;1%SL可获得与卵黄相似的保护效果;在含有卵黄的基础冷冻液中添加CLC未能提高精子冻后活力。

In the present study,we investigated the effect of different yolk components（whole egg yolk（EY）,egg yolk plasma proteins（EYP）,low-density lipoprotein（LDL）and livetin）,soy lecithin（SL）and cholesterol-loaded cyclodextrin（CLC）on equine sperm cryopreservation.Semen was collected from 4adult thoroughbred stallions,experiments 1：Adding 0,2%,4%and 8%concentration of EY,LDL,EYP or livetin in INRA82＋3.5% combination cryoprotectants;Experiments 2：Adding 1%,2% and 4% SL with 4% EY as control group;Experiment 3：Adding 0,0.75,1.5and 3mg/mL concentration of CLC.The results showed that EYP,LDL and livetin had a significant cryoprotective function（P〈0.05）,but no significant difference was observed between different components of the yolk and whole egg yolk（P〉0.05）;Addition of 1% SL ob-tained similar cryopreservation result with the control group（4%egg yolk）（P〉0.05）,but postthaw TM,PM and RAP supplemented with 2% and 4% SL group were significantly lower than the control group（P〈0.05）;There was no significant difference between different concentrations of CLC group and the control group in thawed sperm motility（P〉0.05）.In conclusion,EYP,LDL and livetin had a significant cryoprotective function,but no significant difference with EY;Addition of 1% SL could obtain similar cryopreservation result with the 4% EY group;Supplying CLC in extender contain egg yolk could not improve post-thaw equine semen quality.