摘要
旨在研究AMPK激活剂二甲双胍(metformin,Met)和阿卡地新(acadesine,AICAR)对绵羊精液冷冻保存效果的影响。本研究首先在冷冻基础稀释液中分别添加不同浓度(0、100、200、300、400、500μmol·L-1)的Met和AICAR,冷冻解冻后根据精子活力、运动性能和结构完整性指标筛选出最佳的添加浓度(400μmol·L-1 Met、200μmol·L-1 AICAR);然后分别使用不同的冷冻稀释液(对照组:稀释液;Met组:含400μmol·L-1 Met的稀释液;AICAR组:含200μmol·L-1 AICAR的稀释液)冷冻精液,解冻后检测精子中AMPK蛋白表达、顶体酶活性、代谢指标、线粒体功能以及抗氧化酶活性。结果表明,稀释液中添加400μmol·L-1 Met和200μmol·L-1 AICAR均可显著提高解冻后精子活力、运动性能及精子结构完整性(P<0.05),其中400μmol·L-1 Met组精子总活力达43.20%,顶体完整率为91%,质膜完整率为46%。与对照组相比,Met组和AICAR组解冻后精子中AMPK磷酸化水平显著升高(P<0.05);顶体酶活性显著提高(P<0.05);丙酮酸水平显著下降(P<0.05),乳酸脱氢酶活性、乳酸以及ATP含量均显著升高(P<0.05);与对照组相比,Met和AICAR组稀释液更有利于维持线粒体膜电位(P<0.05),提高ATP酶(P<0.05)以及抗氧化酶的活性(P<0.05)。添加适当浓度的AMPK激活剂可以提高绵羊精液冷冻保存的效果。
This study aimed to investigate the effects of AMPK activators metformin(Met)and acadesine(AICAR)on sheep semen cryopreservation.Firstly,Met and AICAR with different concentrations(0,100,200,300,400,500μmol·L-1)were added into the frozen diluent.After freezing and thawing,the optimal concentration(400μmol·L-1 Met,200μmol·L-1 AICAR)were selected based on sperm motility,motion performance and membrane integrity;Secondly,the collected semen was froze using different frozen diluents(control group:diluent;Met group:diluent+400μmol·L-1 Met;AICAR group:diluent+200μmol·L-1 AICAR).After thawing,the sperm AMPK protein expression,acrosomal enzyme activity,metabolic index,mitochondrial function and antioxidant enzyme activity were examined.The results showed that the addition of 400μmol·L-1 Met and 200μmol·L-1 AICAR could significantly improve sperm motility,motion performance and membrane integrity(P<0.05)after thawing.In 400μmol·L-1 Met group,the total sperm motility,acrosome integrity rate and plasma membrane integrity rate were 43.20%,91%and 46%,respectively.Compared with the control group,the level of AMPK phosphorylation in the sperm of the Met and AICAR groups was significantly increased after thawing(P<0.05),the acrosome enzyme activity of sperm was significantly increased(P<0.05);the pyruvate level was significantly decreased(P<0.05),the lactate dehydrogenase activity,lactic acid content,and ATP content were significantly increased(P<0.05).Compared with the control group,the dilution of Met group and AICAR group was more conducive to maintain mitochondrial membrane potential(P<0.05),increase ATPase and antioxidant enzyme activity(P<0.05).The addition of appropriate concentrations of Met and AICAR could improve semen quality of cryopreservation in sheep.
作者
汪棋
汪长建
魏宗友
陆汉希
姚晓磊
杨花
王锋
张艳丽
WANG Qi;WANG Changjian;WEI Zongyou;LU Hanxi;YAO Xiaolei;YANG Hua;WANG Feng;ZHANG Yanli(Jiangsu Livestock Embryo Engineering Laboratory,College of Animal Science and Technology,Nanjing Agricultural University,Nanjing 210095,China;Taicang Animal Husbandry and Veterinary Station,Taicang 215400,China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2020年第12期3033-3045,共13页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
南京农业大学动科学院青年人才拔尖项目(DKQB201901)
江苏现代农业产业技术体系项目(JATS[2019]128,JATS[2020]427)。
作者简介
汪棋(1995-),女,安徽安庆人,硕士生,主要从事动物生殖调控研究;E-mail:2018805121@njau.edu.cn;通信作者:张艳丽,主要从事肉羊繁殖调控研究,E-mail:zhangyanli@njau.edu.cn。
