褪黑素改善绵羊冷冻卵母细胞损伤与发育潜能的研究  

Melatonin alleviates the damage and improves the developmental potential of frozen ovine oocytes

作　　者：闫嘉欣 刘炳男 高子浩 张琳惠 李嘉 张慈 陶晨雨 李俊杰[1,2] 夏威 YAN Jiaxin;LIU Bingnan;GAO Zihao;ZHANG Linhui;LI Jia;ZHANG Ci;TAO Chenyu;LI Junjie;XIA Wei(College of Animal Science and Technology,Hebei Agricultural University,Baoding 071000,China;Hebei Province Technology Innovation Center of Cattle and Sheep Embryo,Baoding 071000,China)

机构地区：[1]河北农业大学动物科技学院,河北保定071000 [2]河北省牛羊胚胎技术创新中心,河北保定071000

出　　处：《畜牧与兽医》2025年第4期8-13,共6页Animal Husbandry & Veterinary Medicine

基　　金：国家重点研发计划项目(2021YFD1200401);河北省自然科学基金联合基金项目(C2023204007);河北省教育厅重点项目(ZD2022005);河北省留学回国人才项目(ZD20220513);河北农业大学人才引进专项项目(YJ2021013)。

摘　　要：为研究褪黑素(MLT)对绵羊GV期卵母细胞玻璃化冷冻效果的影响,选取具有3层以上致密完整卵丘细胞且胞质均匀的卵丘-卵母细胞复合体,在玻璃化冷冻解冻液中添加不同浓度(0、10^(-11)、10^(-9)、10^(-7)、10^(-5)mol/L)MLT,解冻后检测体外成熟液培养24 h后卵母细胞以及新鲜组卵母细胞活性氧(ROS)、Ca^(2+)、线粒体膜电位(MMP)水平。结果:与新鲜组相比,0 mol/L MLT组ROS、Ca^(2+)水平显著升高(P<0.05),MMP水平显著降低(P<0.05);相较于0 mol/L MLT组,在玻璃化冷冻解冻液中添加MLT可显著降低绵羊玻璃化卵母细胞ROS和Ca^(2+)水平(P<0.05),并显著提升卵母细胞MMP水平(P<0.05),其中绵羊GV期卵母细胞玻璃化冷冻过程中MLT的最适添加浓度为10-9mol/L;与0 mol/L MLT组相比,10^(-9)mol/L组可显著提高卵母细胞体外受精后卵裂率(68.64%vs 58.03%,P<0.05)与囊胚率(18.97%vs 5.96%,P<0.05)。本研究证实在玻璃化冷冻解冻液中补充MLT可缓解绵羊GV期卵母细胞冷冻损伤并提高其发育潜能。In order to determine the effect of melatonin(MLT)on the vitrification freezing of GV ovine oocytes,we selected the cumulus oocyte complex with more than three layers of dense intact cumulus cells and uniform cytoplasm,and added different concentrations(0,10^(-11),10^(-9),10^(-7),10^(-5)mol/L)of melatonin to the vitrification freezing and thawing solution.After thawing,the levels of reactive oxygen species(ROS),Ca^(2+)and mitochondrial membrane potential(MMP)of oocytes cultured in vitro maturation medium for 24 hours were detec⁃ted.The results showed that,compared with the fresh group,the levels of ROS and Ca^(2+)in the 0 mol/L MLT group were significantly in⁃creased(P<0.05),and the level of MMP was significantly decreased(P<0.05).Compared with the 0 mol/L MLT group,the addition of melatonin in vitrification freezing and thawing solution significantly reduced the ROS level and Ca^(2+)concentration(P<0.05),and signifi⁃cantly increased the MMP level of the oocytes(P<0.05).The optimal melatonin concentration was 10-9 mol/L during vitrification of the GV stage oocytes.Compared with the 0 mol/L MLT group,the 10^(-9) mol/L group significantly increased the cleavage rate of oocytes(68.64%vs 58.03%,P<0.05)and the blastocyst rate(18.97%vs 5.96%,P<0.05).This study confirmed that melatonin supplementation in vitrifica⁃tion freezing and thawing solution alleviated GV stage ovine oocytes cryoinjury and improved their developmental potential.

关 键 词：绵羊 卵母细胞 褪黑素 玻璃化 

分 类 号：S814.8[农业科学—畜牧学]