抗冷冻蛋白Ⅲ对玻璃化冷冻猪GV期卵母细胞及其DNA的保护作用  

Protection of antifreeze protein Ⅲ on vitrified GV oocytes and its DNA of porcine

作　　者：董胤余 王勇杰 刘可可 王欣雨 唐非台 杜倩笙 王晓丽[1] DONG Yin-yu;WANG Yong-jie;LIU Ke-ke;WANG Xin-yu;TANG Fei-tai;DU Qian-sheng;WANG Xiao-li(College of Animal Science and Technology,Guangxi University,Nanning 530005,China)

机构地区：[1]广西大学动物科学技术学院,广西南宁530005

出　　处：《中国兽医科学》2023年第5期658-663,共6页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31460644);广西自然科学基金项目(2018GXNSFAA281020)。

摘　　要：为了寻找冷冻液中最适合的抗冷冻蛋白Ⅲ(AFPⅢ)添加浓度,并探索AFPⅢ对玻璃化冷冻卵母细胞、活性氧(ROS)及其DNA损伤的保护作用进行评价。本研究分别在冷冻液中添加250、500、750、1000、1200 ng/m L AFPⅢ,通过冷冻液处理和冷冻-复苏筛选出最合适的AFPⅢ添加浓度,荧光探针检测卵母细胞中活性氧的含量,彗星电泳检测卵母细胞DNA的变化。结果显示,冷冻液中添加500 ng/m L AFPⅢ对玻璃化冷冻猪GV期卵母细胞的保护效果最佳;冷冻液中添加AFPⅢ的冷冻组ROS含量显著低于冷冻对照组(P<0.05);添加AFPⅢ的冷冻组尾部DNA百分比(Tail DNA%)和Olive尾矩值(OTM)显著低于冷冻对照组(P<0.05)。结果表明,AFPⅢ可以提高玻璃化冷冻后卵母细胞的体外发育能力,降低冷冻-复苏后卵母细胞ROS含量,缓解玻璃化冷冻引起的卵母细胞DNA损伤。In order to explore the most appropriate concentration of antifreeze protein Ⅲ(AFP Ⅲ)in the freezing solution,and to explore the protection of AFP Ⅲ on vitrified GV oocytes of porcine,reactive oxygen species(ROS) content and DNA damage.250,500,750,1000 and 1200 ng/m L AFP Ⅲ were added to the freezing solution respectively,and the most appropriate concentration of AFP Ⅲ was selected through the treatment of freezing solution and freeze-recovery.The content of ROS in oocytes was detected by fluorescence probe.The changes of oocyte DNA were detected by comet electrophoresis.The results showed that 500 ng/m L AFP Ⅲ had the best protective effect on vitrified oocytes at GV stage of porcine.The content of ROS in the AFP Ⅲ addition group was significantly lower than that in the frozen control group(P<0.05).The tail DNA% and olive tail moments(OTM) of AFP Ⅲ addition group were significantly lower than those of frozen control group(P<0.05).The results indicated that AFP Ⅲ could improve the in vitro development ability and reduce the ROS content after freeze-recovery,and alleviate the DNA damage caused by vitrification of GV oocytes in porcine.

关 键 词：抗冷冻蛋白Ⅲ 卵母细胞 玻璃化冷冻 活性氧 DNA损伤 猪 

分 类 号：S828.34[农业科学—畜牧学]