稀释液添加不同抗氧化剂对冷冻姜曲海猪精子质量的影响  

Protective effects of three different antioxidants on frozen sperm in Jiangquhai boar

作　　者：王琳琳[1] 李乐康 戴乾 魏春来 孙雨迪 张婉蓉 谭菊[1] 武彩红[1] WANG Linlin;LI Lekang;DAI Qian;WEI Chunlai;SUN Yudi;ZHANG Wanrong;TAN Ju;WU Caihong(Jiangsu Agri-animal Husbandry Vocational College,Taizhou 225300,China)

机构地区：[1]江苏农牧科技职业学院,江苏泰州225300

出　　处：《畜牧与兽医》2024年第11期10-15,共6页Animal Husbandry & Veterinary Medicine

基　　金：2023年江苏省职业院校学生创新创业培育计划项目(G-2023-0727);江苏农牧科技职业学院动物疫病科技创新团队项目(NSF2021TC03);泰州市科技支撑计划(农业)项目(TN201615)。

摘　　要：旨在探究稀释液中添加不同抗氧化剂对冷冻姜曲海猪精子的保护作用。采用精子直线运动检测精子活力,异硫氰荧光素标记的花生凝集素(FITC-PNA)染色检测精子顶体完整性,碘化丙啶(PI)染色检测精子质膜完整性,体外受精和培养检测受精卵的卵裂情况,透射电镜观察精子超微结构,评价冷冻保存对精子质量的影响;对姜曲海猪精液分别冷冻保存1周、1个月、3个月、6个月、9个月和12个月,观察冷冻保存时间对精子质量的影响;在稀释液中分别添加抗氧化剂甲磺酸米托醌(MitoQ),2,4-二硝基酚(DNP)和4-羟基-2,2,6,6-四甲基哌啶-1-烃氧基(Tempol),每种抗氧化剂均添加25、50和100μmol/L 3种浓度,以筛选出冷冻保存效果最优的抗氧化剂;对添加50μmol/L MitoQ的解冻精子进行透射电镜观察。结果显示:精子冷冻保存不同时间后,各指标均无显著差异(P>0.05);添加不同浓度的MitoQ、DNP和Tempol均对冷冻保存1周精子的活力、顶体完整率、质膜完整率和体外受精卵裂率有一定的改善作用,但以添加50μmol/L MitoQ效果最好;添加50μmol/L MitoQ对精子质膜、线粒体和微管等结构均具有一定的冷冻保护作用。综上,稀释液中添加50μmol/L MitoQ对于超低温保存姜曲海猪精液具有潜在的保护优势,具有一定的开发价值。The aim of the study was to investigate the protective effects of different antioxidants on frozen sperm in Jiangquhai boar.To evaluate the effect of cryopreservation on sperm quality,the sperm motility was detected by sperm linear movement,the acrosome integrity was detected by fluorescein isothiocyanin-labeled peanut agglutinin(FITC-PNA)staining,the plasma membrane integrity was detected by propidium iodide(PI)staining,the cleavage of fertilized eggs was detected by in vitro fertilization,and the sperm ultrastructure was observed by transmission electron microscopy(TEM).The semen was cryopreserved for 1 week,1 month,3 months,6 months,9 months and 12 months,respectively,to observe the effects of different storage time lengths on sperm quality.In order to screen out the antioxidants with the best cryopreservation effect,mitoquinone mesylate(MitoQ),2,4-dinitrophenol(DNP)and 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl(Tempol)was supplemented in the diluent at concentrations of 25,50 and 100μmol/L,respectively.The thawed sperm with 50μmol/L MitoQ were observed by TEM.The results showed that there was no significant difference in all parameters after sperm cryopreservation for different time periods(P>0.05).The addition of MitoQ,DNP and Tempol at different concentrations improved sperm motility,acrosome integrity,plasma membrane integrity and in vitro fertilization cleavage rate after 1-week cryopreservation.However,the addition of 50μmol/L MitoQ had the best effect.In addition,TEM showed that the addition of 50μmol/L MitoQ protected the plasma membrane,mitochondria and microtubules of the sperm.In conclusion,the addition of 50μmol/L MitoQ to the diluent presented potential protective advantages in cryopreservation of sperm in Jiangquhai boars,and MitoQ had certain development value.

关 键 词：姜曲海猪 超低温冷冻 抗氧化剂 精子品质 超微结构 

分 类 号：S813.9[农业科学—畜牧学]