不同浓度维生素E对沂蒙黑猪精液冷冻保存效果的影响

Effects of different concentrations of vitamin E on semen cryopreservation in Yimeng black pigs

为了探讨在冷冻稀释液中添加不同浓度维生素E对沂蒙黑猪精液冷冻保存效果的影响,试验将6头沂蒙黑猪的精液混合后分成6组,分别为对照组(冷冻保存液)和在冷冻保存液中添加100,200,300,400,500 mg/L维生素E的试验组,先用液氮保存,然后将每组精液解冻后检测精子活力及运动参数、顶体完整性、质膜完整性、DNA完整性、过氧化氢酶(CAT)活性、总抗氧能力(T-AOC)、丙二醛(MDA)含量、线粒体膜电位(MMP)和活性氧(ROS)水平,以此来评估维生素E对精液品质的影响。结果表明:与对照组相比,200 mg/L组和300 mg/L组精子活力及各项运动参数均显著提高(P<0.05),其中300 mg/L组提高最显著;100 mg/L组和400 mg/L组精子运动参数也提高,而500 mg/L组精子活力及各项运动参数均显著降低(P<0.05)。与对照组相比,100 mg/L组、200 mg/L组和300 mg/L组精子质膜完整性、顶体完整性、DNA完整性及MMP水平均显著提高(P<0.05);而500 mg/L组精子质膜完整性、顶体完整性及MMP水平均显著降低(P<0.05)。除500 mg/L组精子T-AOC、CAT活性、MDA含量和ROS水平与对照组相比差异不显著(P>0.05)外,其余浓度组与对照组相比,精子T-AOC、CAT活性均显著提高(P<0.05),MDA含量均显著降低(P<0.05)。说明在精液冷冻过程中添加维生素E对沂蒙黑猪精子的保护作用呈浓度依赖性,较低浓度的维生素E对冷冻保存的精子活力有一定的改善作用,而较高浓度的维生素E对冷冻保存的精子活力、质膜完整性、顶体完整性及DNA完整性有显著的不利影响,但不影响精子氧化应激状态。

In order to investigate the effects of adding different concentrations of vitamin E on semen cryopreservation of Yimeng black pigs,the semens of 6 Yimeng black pigs were mixed and divided into 6 groups,the control group(cryopreservation solution)and the experimental groups with 100,200,300,400,500 mg/L of vitamin E added to the cryopreservation solution,respectively,which were treated with liquid nitrogen storage.Then,after thawing each group of semen,sperm motility and motility parameters,acrosome integrity,plasma membrane integrity,DNA integrity,catalase(CAT)activity,total antioxidant capacity(T-AOC),malondialdehyde(MDA)content,mitochondrial membrane potential(MMP)and reactive oxygen species(ROS)levels were detected,which was used to assess the effects of vitamin E on semen quality.The results showed that compared with the control group,sperm motility and motility parameters were significantly improved in the 200 mg/L group and the 300 mg/L group(P<0.05),and the 300 mg/L group had the most significant improvement.The motility parameters of sperm in the 100 mg/L group and the 400 mg/L group were also improved,while the motility and motility parameters in the 500 mg/L group were significantly reduced(P<0.05).Compared with the control group,the sperm plasma membrane,acrosome,DNA integrity rate and MMP levels were significantly increased in the 100 mg/L group,200 mg/L group and 300 mg/L group(P<0.05),while the sperm plasma membrane,acrosome integrity rate and MMP level in the 500 mg/L group were significantly reduced(P<0.05).Except for the 500 mg/L group in which the sperm T-AOC,CAT activity,MDA content and ROS level were not significantly different from the control group(P>0.05),and the sperm T-AOC and CAT activity were significantly increased(P<0.05),and MDA content were significantly decreased(P<0.05)in other dose groups compared with the control group.The results suggested that the protective effect of adding vitamin E on the sperm of Yimeng black pigs during semen freezing was concentration-dependent.Low concentration of vitamin E had some improvement on the viability of cryopreserved sperm.In contrast,high concentration of vitamin E had significant adverse effects on the viability,plasma membrane,acrosome and DNA integrity of cryopreserved sperm,but did not affect the oxidative stress state of sperm.