摘要
为探究高压均质鸡蛋卵黄对猪冷冻精子凋亡的影响,本试验采集5头健康状态良好的杜洛克公猪精液,以普通鸡蛋卵黄+Tris-柠檬酸-葡萄糖(TCG)冷冻基础稀释液为对照组,以添加高压均质处理的鸡蛋卵黄+Tris-柠檬酸-葡萄糖(TCG)冷冻基础稀释液为试验组;精液冷冻-解冻后观察精子活力、DNA完整性、线粒体膜电位变化、细胞凋亡率、多种Caspase活性;同时利用qRT-PCR检测相关凋亡功能基因的mRNA表达水平,并进行数据统计。结果表明,经高压均质处理后,猪精子冻后活力、活率和顶体完整率显著高于对照组(P<0.05),为87.64%、87.14%和66.61%,较对照组分别提高了12.58%、5.82%和12.48%;线粒体膜电位和DNA完整率显著高于对照组(P<0.05),为0.74和61.76%;精子凋亡水平显著减少(P<0.05),为37.74%;试验组的Caspase-3、Caspase-8和Caspase-9活性为17.15、11.19和15.18,较对照组分别减少了6.17、2.18和3.51(P<0.05);对照组的Bax、TNF-α、Caspase-9基因表达均明显高于试验组(P<0.05),Bcl-2基因表达量较试验组降低(P>0.05)。综上,高压均质鸡蛋卵黄能提高猪精子冻后质量,促进线粒体功能完整性、降低精子细胞早期凋亡水平和细胞内Caspase活性,同时降低凋亡相关基因mRNA表达量。
The aim of this study was to explore the effect of egg yolk(EY)treated by high pressure homogenization on the freezing effect of boar semen.Semen were collected from 5 healthy,mature Duroc boars.The ordinary egg yolk+Tris-citric acid-glucose(TCG)diluent was the control group,and the high pressure homogenized egg yolk+Tris-citric acid-glucose(TCG)diluent was the treatment group.Frozen semen samples were thawed,and the sperm motility,DNA integrity,changes in mitochondrial membrane potential,number of apoptotic cells,Caspases activity were detected.The expression of apoptosis-related genes mRNA was measured by qRT-PCR,and extracting raw data and performing data were analyzed.The results showed that,after the high pressure homogenization treatment,the vitality,motility and acrosomal integrity of pig sperms after freezing were significantly higher than those of the control group(P<0.05),which were 87.64%,87.14%and 66.61%,respectively,higher than those of the control group for 12.58%,5.82%and 12.48%;mitochondrial membrane potential and DNA integrity were significantly higher than those of the control group(P<0.05),which were 0.74 and 61.76%;the level of sperm apoptosis was significantly reduced(P<0.05),which was 37.74%;the Caspase-3,Caspase-8 and Caspase-9 activities of the experimental group were 17.15,11.19 and 15.18,which were reduced by 6.17,2.18 and 3.51 compared with the control group(P<0.05);the expression of Bax,TNF-α,and Caspase-9 genes in the control group were significantly higher than those in the treatment group(P<0.05),and the Bcl-2 gene expression was lower than that in the treatment group(P>0.05).In summary,high pressure homogenous egg yolk can improve the quality of pig sperms after freezing,promote the integrity of mitochondrial function,reduce the level of early sperm cell apoptosis and intracellular Caspase activity,and reduce the mRNA expression of apoptosis-related genes.
作者
汪俊跃
戴建军
张树山
孙玲伟
吴彩凤
张德福
WANG Junyue;DAI Jianjun;ZHANG Shushan;SUN Lingwei;WU Caifeng;ZHANG Defu(Division of Animal Genetic Engineering, Shanghai Municipal Key Laboratory of Agri-genetics and Breeding,Institute of Animal Science and Veterinary Medicine, Shanghai Academy of Agricultural Sciences, Shanghai 201106,China;National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306,China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2021年第8期2190-2199,共10页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
上海市科委“科技创新行动计划”(18391903700)
科技创新2025重大专项(2019B10023)。
关键词
高压均质
鸡蛋卵黄
精子冷冻
细胞凋亡
high pressure homogenization
egg yolk
semen cryopreservation
cell apoptosis
