褪黑色素对兔精子冷冻保存效果的影响
发布时间：2018-11-14 08:40
【摘要】：兔精子在冷冻保存的过程中,由于遭受冷休克而产生过量的对精子有害的氧自由基,致使精子容易发生脂质过氧化反应,从而使精子的质膜遭到破坏,导致解冻后的精子品质降低、人工授精率、分娩率降低,限制了兔人工授精技术的推广运用。本试验拟在兔精子冷冻保存液TCG中添加不同浓度的褪黑色素,初步探讨褪黑色素对兔精子冷冻保存的影响,利用光学显微镜观察精子活率,且利用SYBR14/PI、FITC-PNA、JC-1、BODIPY581/591C11荧光染料检测解冻后的精子的质膜完整率、顶体完整率、线粒体膜电位变化及脂质过氧化反应,此外,还对褪黑色素处理的冻存后的精子进行凋亡的检测,研究获得以下结果:1.在兔精子冷冻保存稀释液中,添加不同浓度的褪黑色素,与对照组相比,可以显著提高解冻后的兔精子的活率、质膜完整率、顶体完整率、线粒体膜电位变化及降低了精子的脂质过氧化反应(P0.05),且褪黑色素的推荐添加浓度为0.1或1mM,当添加浓度为5mM时,效果不显著(P0.05)。2.在精子冷冻稀释液中,添加褪黑色素可以减缓精子因在冷冻-解冻过程中遭受损伤而引起的凋亡现象。试验发现,当在稀释液中添加1mM的褪黑色素时,精子的凋亡现象可以得到缓解。因此,在兔精子冷冻保存中,添加0.1mM的褪黑色素可能通过减缓精子的凋亡,从而提高解冻后的精子品质。
[Abstract]:During the cryopreservation of rabbit spermatozoa, excessive oxygen free radicals harmful to sperm are produced due to cold shock, which makes the sperm prone to lipid peroxidation, thus destroying the plasma membrane of the sperm. After thawing, the sperm quality, artificial insemination rate and delivery rate decreased, which limited the application of artificial insemination technology in rabbits. In this experiment, different concentrations of melatonin were added to rabbit sperm cryopreservation solution (TCG). The effect of melatonin on rabbit sperm cryopreservation was studied. The sperm motility was observed by optical microscope and SYBR14/PI,FITC-PNA,JC-1, was used. The plasma membrane integrity rate, acrosome integrity rate, mitochondrial membrane potential change and lipid peroxidation of thawed spermatozoa were detected by BODIPY581/591C11 fluorescent dye. In addition, apoptosis of frozen spermatozoa treated with melatonin was also detected. The results are as follows: 1. Compared with the control group, the sperm motility, plasma membrane integrity and acrosome integrity of rabbit spermatozoa were significantly increased by adding melatonin at different concentrations in the rabbit sperm cryopreservation diluent. The change of mitochondrial membrane potential and the decrease of lipid peroxidation of spermatozoa (P0.05), and the recommended concentration of melatonin was 0.1 or 1 mm, but the effect was not significant when the concentration of 5mM was added (P0.05). The addition of melatonin in semen cryopreservation diluents can attenuate the apoptosis of spermatozoa caused by injury during freezing and thawing. It was found that the apoptosis of spermatozoa could be alleviated when 1mM melatonin was added to the diluent. Therefore, in cryopreservation of rabbit spermatozoa, melatonin added with 0.1mM may improve the quality of thawed spermatozoa by slowing down the apoptosis of spermatozoa.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S829.1
本文编号：2330681
[Abstract]:During the cryopreservation of rabbit spermatozoa, excessive oxygen free radicals harmful to sperm are produced due to cold shock, which makes the sperm prone to lipid peroxidation, thus destroying the plasma membrane of the sperm. After thawing, the sperm quality, artificial insemination rate and delivery rate decreased, which limited the application of artificial insemination technology in rabbits. In this experiment, different concentrations of melatonin were added to rabbit sperm cryopreservation solution (TCG). The effect of melatonin on rabbit sperm cryopreservation was studied. The sperm motility was observed by optical microscope and SYBR14/PI,FITC-PNA,JC-1, was used. The plasma membrane integrity rate, acrosome integrity rate, mitochondrial membrane potential change and lipid peroxidation of thawed spermatozoa were detected by BODIPY581/591C11 fluorescent dye. In addition, apoptosis of frozen spermatozoa treated with melatonin was also detected. The results are as follows: 1. Compared with the control group, the sperm motility, plasma membrane integrity and acrosome integrity of rabbit spermatozoa were significantly increased by adding melatonin at different concentrations in the rabbit sperm cryopreservation diluent. The change of mitochondrial membrane potential and the decrease of lipid peroxidation of spermatozoa (P0.05), and the recommended concentration of melatonin was 0.1 or 1 mm, but the effect was not significant when the concentration of 5mM was added (P0.05). The addition of melatonin in semen cryopreservation diluents can attenuate the apoptosis of spermatozoa caused by injury during freezing and thawing. It was found that the apoptosis of spermatozoa could be alleviated when 1mM melatonin was added to the diluent. Therefore, in cryopreservation of rabbit spermatozoa, melatonin added with 0.1mM may improve the quality of thawed spermatozoa by slowing down the apoptosis of spermatozoa.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S829.1
【参考文献】
相关期刊论文 前3条
1 马忠兰;不同稀释液对猪精液保存效果的研究[J];黑龙江畜牧兽医;2004年04期
2 杜立银;曹少先;郭炜;王海英;刘铁铮;;褪黑素在猪精液保存中的抗氧化作用[J];江苏农业学报;2009年02期
3 柴局;高日明;张家新;孟克格日乐;王惠梅;赵艳红;王志新;张文广;李金泉;;JC-1单标法和JC-1/PI双标法荧光探针检测山羊附睾精子线粒体膜功能的研究[J];湖北农业科学;2011年12期
相关硕士学位论文 前1条
1 胡靓华;褐藻酸钠、甜菜碱和褪黑素对猪精子冷冻保存效果的影响[D];西北农林科技大学;2014年
，本文编号：2330681
本文链接：https://www.wllwen.com/yixuelunwen/dongwuyixue/2330681.html
