维甲酸对体外热处理猪精子活力、抗氧化性能及其糖脂代谢的影响  

Effect of retinoic acid on motility,antioxidant capacity and glycolipid metabolism of heat-treated boar sperm in vitro

作　　者：石一凡 李晓彤 刘俊泽 李春梅[1] 李延森[1] SHI Yifan;LI Xiaotong;LIU Junze;LI Chunmei;LI Yansen(Research Center for Livestock Environmental Control and Smart Production,College of Animal Science and Technology,Nanjing Agriculture University,Nanjing 210095,China)

机构地区：[1]南京农业大学动物科技学院家畜环境控制与智慧生产研究中心,江苏南京210095

出　　处：《畜牧与兽医》2023年第11期1-10,共10页Animal Husbandry & Veterinary Medicine

基　　金：中国博士后科学基金项目(2020M681648);江苏省博士后科研资助计划项目(2021K435C);江苏现代农业(生猪)产业技术体系建设项目(JATS[2022]083)。

摘　　要：旨在探究维甲酸(RA)对体外热处理猪精子活力、抗氧化性能及其糖脂代谢的影响。将相同来源的精液离心重悬于EBSS培养基,随机分为5组,在39℃环境条件下分别孵育0、1、2、3和4 h后,测定精子活力,筛选合适的孵育时间。将相同来源精液离心重悬后,随机分为6组:对照(CON)组、10、50、100、1000 nmol·L^(-1)RA组和DMSO组,在39℃环境条件下孵育3 h,测定精子活力,筛选RA适宜添加剂量。将相同来源精液离心重悬后,随机分为4组:37℃、37℃+RA、39℃、39℃+RA。37℃+RA组和39℃+RA组培养基中RA浓度为100 nmol·L^(-1),37℃组和39℃组培养基中不含RA,37℃组和37℃+RA组于37℃环境下孵育,39℃组和39℃+RA组于39℃下孵育,测定精子活力,并利用蛋白荧光免疫方法,检测维甲酸受体α(RARα)蛋白定位表达,收集精子用于精子抗氧化和糖脂代谢指标的测定。结果显示:1)39℃处理条件下,随着孵育时间的增加,精子的运动性能逐渐变差。2)39℃处理3 h条件下,RA能改善精子运动性能,且浓度为100 nmol·L^(-1)时效果最好。3)与37℃组相比,39℃培养会显著降低精子总活力及前进运动精子比例(P<0.05),显著提高不运动精子比例(P<0.05);与39℃组相比,培养基中含100 nmol·L^(-1)RA能显著提高精子总活力、前进运动精子比例以及快速运动精子比例(P<0.05),显著降低不运动精子比例(P<0.05);与37℃组相比,39℃培养能显著增加精子丙二醛(MDA)水平(P<0.05),降低T-SOD活力(P<0.05);与39℃组相比,培养基中含100 nmol·L^(-1)RA能显著降低精子MDA水平(P<0.05),提高精子T-SOD活力(P<0.05)。39℃培养和RA处理对精子丙酮酸、葡萄糖、总胆固醇及甘油三酯含量无显著影响。4)RARα蛋白表达主要集中在精子尾部的主段和末段,头部和富含线粒体的中段有较弱表达,但不明显。100 nmol·L^(-1)RA未明显改变RARα蛋白表达位置。结论:猪精子能够表达RARα蛋白,热处理使精子�The aim of this experiment was to investigate the effects of retinoic acid(RA)on motility,antioxidant capacity and glycolipid metabolism of heat-treated boar sperm in vitro.Three experiments were designed in this paper.Firstly,semen samples from the same source were centrifuged and suspended in EBSS medium and randomly divided into 5 groups.After incubation at 39℃for 0,1,2,3 and 4 h,sperm motility of the samples was measured and the appropriate incubation time was selected.Secondly,semen samples from the same source were centrifuged and suspended,and randomly divided into 6 groups:the CON group,the 10,50,100,1000 nmol·L-1 RA groups and the DMSO group,and were incubated at 39℃for 3 h to determine sperm motility and select the appropriate dosage of RA.Finally,semen samples from the same source were centrifuged and suspended,and randomly divided into 4 groups:the 37℃,37℃+RA,39℃and 39℃+RA groups.The concentrations of RA in the 37℃+RA group and the 39℃+RA group were 100 nmol·L-1,and the culture medium of the 37℃and 39℃groups did not contain RA,the 37℃and 37℃+RA groups were incubated at 37℃,and the 39℃and 39℃+RA groups were incubated at 39℃.Sperm motility was measured.The localized expression of retinoic acid receptorα(RARα)protein was de⁃tected by protein fluorescence immunoassay,and sperm samples were collected to determine the antioxidant and metabolic indexes of the sperms.The results showed as follows:1)After cultured at 39℃in vitro,the motility of the sperms gradually deteriorated with the increase of incubation time.2)RA could improve sperm motility at 39℃for 3 h,and the best effect was achieved at 100 nmol·L-1.3)Compared with the 37℃group,the total sperm motility and the rate of progressive motile sperms were significantly decreased when cultured at 39℃(P<0.05),and the rate of immotile sperms was significantly increased(P<0.05).Compared with the 39℃group,the total motility,the rate of progressive motile sperms and the rate of fast motile sperms in the 39℃+RA gr

关 键 词：维甲酸 猪精子 热处理 抗氧化 糖脂代谢 维甲酸受体Α 

分 类 号：S828[农业科学—畜牧学]