巴马香猪精液冷冻过程中精子内抗氧化酶活性与精子质膜完整性关系研究  

The Relationship Between Intracellular Antioxidant Enzyme Activity and Sperm Cell Membrane Integrity in Bama Miniature Pig Semen Cryopreservation

作　　者：孔德营[1] 商海涛[2] 刘福慧[3] 韩勇[1] 魏泓[2] 张家骅[4] 

机构地区：[1]遵义医学院基础医学院,贵州遵义563000 [2]第三军医大学基础部,重庆400038 [3]遵义医学院附属医院输血科,贵州遵义563000 [4]西南大学动物科技学院/重庆市草食动物资源保护与利用工程技术研究中心,重庆400716

出　　处：《西南大学学报（自然科学版）》2014年第9期23-28,共6页Journal of Southwest University(Natural Science Edition)

基　　金：国家重点基础研究发展计划(973计划)重大科学问题导向项目资助(2011CBA01006);遵义医学院重点学科建设项目资助(XZXK-20120702);2013年重庆高校创新团队建设计划资助

摘　　要：猪精液冷冻过程中,精子质膜往往因受到活性氧(ROS)的氧化而发生损伤,精子品质也随之下降.精子ROS水平受抗氧化酶活性的影响,然而冷冻过程中精子细胞内抗氧化酶活性变化与细胞膜完整性的关系还缺乏报道.该试验将巴马香猪精液冷冻保存降温过程中的精子分为4种状态,即鲜精、15℃精液、5℃精液和冷冻-解冻后精液,检测了各种状态下精子的运动参数、质膜完整性(PMI)和顶体完整性(AI),分析了超氧化物歧化酶(SOD)活性、谷胱甘肽过氧化物酶(Gpx)活性和ROS(丙二醛-MDA)水平.结果表明:15℃精子各项指标和鲜精无显著差异(p>0.05);5℃精子运动能力、质膜完整性和抗氧化酶活性低于鲜精(p<0.05),而精子细胞内MDA水平与鲜精无显著差异(p>0.05);冷冻-解冻后精子的运动能力、质膜完整性和抗氧化酶活性均低于其他各组(p<0.05),精子MDA水平显著高于其他各组(p<0.05).同时,SOD和Gpx活性与精子PMI和AI之间的相关性均达到了显著水平(p<0.05).可见,当精子降温至15℃以下直至冷冻时,SOD和GPX活性逐渐降低,精子细胞清除ROS的能力随之下降,精子质膜脂质过氧化反应增强,质膜受损,内容物外渗是精液品质降低的重要原因.During Bama miniature pig semen cryopreservation ,Sperm cell membrane can be damaged due to the oxidation of ROS ,and thus affect Sperm quality .The level of ROS in sperm cells affected by the activities of antioxidant enzymes ,however ,no data are available concerning the relationship between sperm intracellular antioxidant enzyme activity and cell membrane integrity during semen cryopreservation .In our experiment ,we divided the sperm during cryopreservation process into four statuses ,such as fresh sperm , sperm at 15 ℃ ,sperm at 5 ℃ ,and frozen-thawed sperm ,and detected the motion parameters ,plasma membrane integrity （PMI） ,acrosome integrity （AI） of sperm in these statuses ,analysed the activity of superoxide dismutase （SOD） ,glutathione peroxidase （Gpx） and the level of ROS （malonaldehyde-MDA） in sperm cells .Results showed that ,sperm at 15 ℃ and fresh sperm indexes had no significant difference （p〉0.05） .Motility ,cell membrane integrity and antioxidant enzymes activity of sperm at 5 ℃ were low-er than fresh sperm （p 〈 0.05） ,however ,sperm cells MDA levels had no significant differences （p 〉0.05） .Motility ,cell membrane integrity and antioxidant enzyme activities of frozen-thawed sperm were lower than those in other groups （p〈0.05） ,and sperm MDA level is significantly higher than that of other groups （p〈0.05） .Meanwhile ,the correlation between SOD ,Gpx activity and sperm PMI ,AI were significant （p 〈 0.05） .And we can conclude that ,When the temperature was below 15 ℃ to freezing , SOD and GPX activity of sperm decreased gradually ,cell membrane lipid peroxidation of sperm enhanced , cell membrane damaged ,the contents of sperm exosmosed ,and this is an important reason leading to the reduction of semen quality .

关 键 词：巴马香猪 精液冷冻 活性氧 抗氧化酶活性 细胞膜完整性 

分 类 号：S828[农业科学—畜牧学]