当归多糖对崇明白山羊冷冻精液质量与DNA甲基化的影响  

Effects of angelica polysaccharides on frozen sperm quality and DNA methylation in Chongming White goats

作　　者：张克勤 周金勇 何孟纤 张德福[2] 戴建军[2] 张维娅 孙玲伟 ZHANG Keqin;ZHOU Jinyong;HE Mengqian;ZHANG Defu;DAI Jianjun;ZHANG Weiya;SUN Lingwei(College of Animal Science and Technology,Hebei Agricultural University,Baoding 071001,China;Key Laboratory of Agricultural Genetic Breeding Institute of Animal Husbandry and Veterinary Medicine,Shanghai Academy of Agricultural Sciences,Shanghai 201106,China)

机构地区：[1]河北农业大学动物科技学院,河北保定071001 [2]上海市农业科学院畜牧兽医研究所农业遗传育种重点实验室,上海201106

出　　处：《黑龙江畜牧兽医》2024年第15期36-42,117,共8页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金青年基金项目(32202640);上海市农业科学院助跑计划项目(ZP22171)。

摘　　要：为了解决崇明白山羊精液冷冻后总体质量偏低的问题,试验采集8只24~26月龄体型匀称的崇明白山羊精液,检测合格后混匀以消除个体差异,分别用未添加当归多糖(ASP)和添加150,300,600,1200μg/mL ASP的基础稀释液稀释后进行冷冻,解冻后评估精液的质量参数(活率、活力、质膜完整率、顶体完整率、线粒体活性)、相关抗氧化指标[丙二醛(MDA)、活性氧(ROS)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GPH-Px)、一氧化氮(NO)、总抗氧化能力(T-AOC)、过氧化氢酶(CAT)]及DNA甲基化5-MC水平,最终确定ASP的最适宜添加浓度,并分析了精液DNA甲基化5-MC水平与精液质量参数的相关性。结果表明:与未添加ASP相比,添加600μg/mL ASP的精子活率、活力、质膜完整率和线粒体活性都显著升高(P<0.05);添加300μg/mL ASP的精子顶体完整率显著升高(P<0.05);添加600μg/mL ASP的精子SOD、CAT活性及T-AOC水平都显著升高(P<0.05);添加600μg/mL ASP的精子ROS、MDA含量都显著降低(P<0.05);添加300,600μg/mL ASP的精子DNA甲基化5-MC水平显著降低(P<0.05)。精液DNA甲基化5-MC水平与精子活力、活率、质膜完整率、顶体完整率、线粒体活性均呈现极显著负相关(P<0.01),R分别为-0.983,-0.892,-0.992,-0.732,-0.825。说明在本试验条件下,基础稀释液中添加600μg/mL ASP最利于崇明白山羊精液的冷冻保存。In order to solve the problem of the diminished quality in the frozen sperm of Chongming white goats,semen samples from eight well-proportioned Chongming white goats aged 24 to 26 months were obtained and mixed to eliminate individual differences after qualified detection.Semen was programmed to freeze after treatment with basal diluent without angelica polysaccharide(ASP)or with 150,300,600,and 1200μg/mL ASP.Sperm quality parameters(viability,motility,plasma membrane integrity,acrosome integrity,mitochondrial activity),relevant oxidative indices(malondialdehyde(MDA),reactive oxygen species(ROS),superoxide dismutase(SOD),glutathione peroxidase(GPH-Px),nitric oxide(NO)content,total antioxidant capacity(T-AOC),catalase(CAT)),and sperm DNA methylation level were evaluated;the optimal concentration of ASP was determined,and the correlation between sperm DNA methylation 5-MC level and sperm quality parameters was analyzed.The results showed that viability,motility,plasma membrane integrity and mitochondrial activity of sperm in the 600μg/mL ASP group were significantly higher than those in the group without ASP(P<0.05),and the acrosome integrity of sperm in the 300μg/mL ASP group was significantly higher than that in the group without ASP(P<0.05).The SOD and CAT activities and T-AOC values of sperm in the 600μg/mL ASP group were significant higher than those in the group without ASP(P<0.05),but ROS and MDA values of sperm in the 600μg/mL ASP group were significantly lower than those in the group without ASP(P<0.05).The DNA methylation level of sperm in the 300 and 600μg/mL ASP groups was significant lower than that in the group without ASP(P<0.05).Sperm DNA methylation 5-MC level was significantly negatively correlated with sperm motility,viability,plasma membrane integrity,acrosome integrity and mitochondrial activity(P<0.01),and R=-0.983,-0.892,-0.992,-0.732,-0.825.The results indicated that the addition of 600μg/mL ASP in basal diluent was the most appropriate for the cryopreservation of Chongming white goat

关 键 词：精液 冷冻保存 崇明白山羊 抗氧化性 当归多糖 DNA甲基化5-MC水平 

分 类 号：S826[农业科学—畜牧学]