白藜芦醇对牛精子体外获能效果及早期胚胎发育的影响  

The Effect of Resveratrol on Sperm Capacitation In vitro and Early Embryo Development in Bovine(Bos taurus)

作　　者：李晓霞[1,2] 曹平华 汪豆豆[1] 曲培栋 徐志谦 禹学礼 栗颖华[1,2] LI Xiao-Xia;CAO Ping-Hua;WANG Dou-Dou;QU Pei-Dong;XU Zhi-Qian;YU Xue-Li;LI Ying-Hua(College of Animal Science and Technology,Henan University of Science and Technology,Luoyang 471023,China;Henan Provincial Key Laboratory for Grass-Feeding Animal,Luoyang 471023,China)

机构地区：[1]河南科技大学动物科技学院,洛阳471023 [2]河南省草食动物工程重点实验室,洛阳471023

出　　处：《农业生物技术学报》2024年第8期1836-1843,共8页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31201801,31872354);湖北省重点实验室开放课题(KLAEMB-2019-06);河南省教育厅重点科研项目计划(19B230004);河南科技大学大学生研究训练计划基金(2023495);“洛阳木叶萱生物科技有限公司”委托保护剂研发项目(横20210029)。

摘　　要：精子体外获能过程中产生的过量活性氧(reactive oxygen species,ROS)会降低精子质量,进而影响早期胚胎发育。白藜芦醇是一种有效保护精子的抗氧化剂。为探讨白藜芦醇对牛(Bos taurus)精子体外获能效果及早期胚胎发育的影响,本研究将解冻后的荷斯坦牛冻精加入到分别含0、10、50、100和500μmol/L白藜芦醇的获能液中,经上游法处理1 h后,评估不同浓度白藜芦醇对牛精子活力、顶体完整性、线粒体活性、质膜完整性、获能状态及体外受精后早期胚胎发育能力的影响。结果表明,与对照组(0μmol/L)相比,在精子获能液中添加50和100μmol/L白藜芦醇的牛精子活力、顶体完整率、线粒体高膜电位百分率和质膜完整率均显著升高(P<0.05),并显著提高了体外获能后B型精子占比率(P<0.05)。金霉素(chlortetracycline,CTC)荧光染色法与考马斯亮蓝染色法均可评估牛精子顶体完整性,两者之间无显著差异(P>0.05)。体外受精结果表明,与对照组(0μmol/L)相比,获能液添加100μmol/L白藜芦醇能显著提高体外受精后的卵裂率和囊胚率(P<0.05)。综上表明,在获能液中添加100μmol/L白藜芦醇能显著改善牛精子功能,提高其获能率,使体外受精后的卵裂率和囊胚率显著提升。本研究有助于优化体外受精体系,加速体外胚胎生产效率。An excess production of reactive oxygen species(ROS)during sperm capacitation in vitro will decrease sperm quality,and affect early embryonic development.Resveratrol is an effective anti-oxidant to protect sperm.In this study,the effect of resveratrol was evaluated on sperm capacitation in vitro and early embryo development in bovine(Bos taurus).The frozen-thawed semen from Holstein bulls was used in sperm capacitation medium supplementing with 0,10,50100 and 500μmol/L resveratrol,subsequently incubated by swim-up method for 1 h.This study was to evaluate the effects of resveratrol on bovine sperm progressive motility,acrosome integrity,mitochondrial activity,plasma membrane integrity,capacitation status and the capacity of early embryo development after in vitro fertilization.These results showed that compared with the control group(0μmol/mL),the supplementation of 50,100μmol/L resveratrol in sperm capacitation medium could significantly increase sperm progressive motility,the percentage of sperm with acrosome integrity,high mitochondrial membrane potential and plasma membrane integrity in bovine after in vitro capacitation(P<0.05);and it significantly increased the percentage of the B pattern sperm.Both of chlortetracycline(CTC)fluorescent staining and coomassie staining could be used to assess acrosome integrity of bovine sperm,and there was no significant difference between the 2 methods(P>0.05).The results of in vitro fertilization showed that compared with the control group(0μmol/mL),the supplementation of resveratrol at 100μmol/L in the capacitation medium resulted in a higher cleavage rate and blastocyst rate(P<0.05).This result showed that adding 100μmol/mL resveratrol to capacitation medium could significantly improve sperm function of bovine after in vitro capacitation,increase its capacitation rate,so that the cleavage and blastocyst rate were significantly increased after in vitro fertilization.This study could help to optimize in vitro fertilization procedures and accelerate the efficiency of

关 键 词：白藜芦醇 牛精子 体外获能 胚胎发育 

分 类 号：S814.8[农业科学—畜牧学]