线粒体靶向抗氧化剂MitoQ在猪卵母细胞冷冻保存中的应用  

Application of mitochondrial targeted antioxidant MitoQ in cryopreservation of porcine oocytes

作　　者：王文杰 葛雷 张树山[2,3] 吴彩风[2,3] 戴建军 张德福[2,3] WANG Wenjie;GE Lei;ZHANG Shushan;WU Caifeng;DAI Jianjun;ZHANG Defu(College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 201306,China;Institute of Animal Husbandry and Veterinary Science,Shanghai Academy of Agricultural Sciences,Shanghai 201106,China;Division of Animal Genetic Engineering,Shanghai Key Laboratory of Agri-Genetics and Breeding,Shanghai 201106,China)

机构地区：[1]上海海洋大学水产与生命学院,上海201306 [2]上海市农业科学院畜牧兽医研究所,上海201106 [3]上海市农业遗传育种重点实验室动物遗传工程研究室,上海201106

出　　处：《上海农业学报》2020年第2期82-88,共7页Acta Agriculturae Shanghai

基　　金：国家自然科学基金(31372315);上海市嘉定区农业和社会事业研究项目(2016004);上海市农业科学院攀高计划(PG171)。

摘　　要：在猪卵母细胞解冻后的孵育液中添加线粒体靶向抗氧化剂MitoQ,通过检测线粒体膜电位、早期凋亡、脂质氧化、活性氧、胚胎发育水平以及相关基因表达水平等,研究MitoQ在猪卵母细胞冷冻保存中的作用.结果表明:猪卵母细胞冷冻后使用MitoQ处理,其线粒体膜电位(0.66±0.06)与对照组(0.47±0.05)相比显著升高,早期凋亡比率(43.00%±3.51%)与对照组(55.33%±1.45%)相比显著下降;冷冻后MitoQ处理猪卵母细胞,其活性氧水平(3.03±0.61)与脂质氧化水平(20.54±0.91)与对照组(13.13±1.52和26.78±0.54)相比显著下降;冷冻后MitoQ处理猪卵母细胞的存活率(60.33%±1.86%)、卵裂率(28.82%±1.38%)和囊胚率(4.50%±0.48%)与对照组组(47.67%±1.67%、22.07%±1.07%和2.20%±0.51%)相比显著升高;在基因表达方面,与对照组相比,冷冻后MitoQ处理组其Bcl-2、CuZnSOD和Mfn2相对表达量显著提升,DNM1、Bax和TNF-α相对表达量则显著下降.综上,在冻后孵育液中添加MitoQ能够提高线粒体功能,降低细胞氧化应激水平,抑制细胞凋亡和改善相关基因表达水平,从而提高冷冻后猪卵母细胞的体外发育能力.MitoQ,a mitochondrial targeted antioxidant,was added to the thawed incubation solution of porcine oocytes.The effects of MitoQ on the cryopreservation of porcine oocytes were studied by detecting mitochondrial membrane potential,early apoptosis,lipid oxidatio,reactive oxygen species(ROS),embryonic development ability and expression level of related genes.The results showed that the mitochondrial membrane potential(0.66±0.06)of porcine oocytes treated with MitoQ was significantly higher than that of the control group(0.47±0.05),and the early apoptosis rate(43.00%±3.51%)was significantly lower than that of the control group(55.33%±1.45%).The levels of ROS(3.03±0.61)and lipid oxidation(20.54±0.91)in the frozen porcine oocytes treated with MitoQ were significantly lower than those in the control group(13.13±1.52,26.78±0.54).The survival rate(60.33%±1.86%),cleavage rate(28.82%±1.38%)and blastocyst rate(4.50%±0.48%)of the porcine oocytes treated with MitoQ were significantly higher than those of the control group(47.67%±1.67%,22.07%±1.07%and 2.20%±0.51%).In terms of gene expression,compared with the control group,the relative expression levels of Bcl-2,CuZnSOD and Mfn2 increased significantly,while the relative expression levels of Dnm1,Bax and TNF-αdecreased significantly in the MitoQ treated group.In summary,the addition of MitoQ in thawed incubation solution could improve the mitochondrial function,reduce the level of oxidative stress,inhibit the apoptosis of cells and improve the expression of related genes,so as to improve the development ability of frozen porcine oocytes in vitro.

关 键 词：猪卵母细胞 Mitoquinone 氧化应激 凋亡 冷冻保护 

分 类 号：S814.8[农业科学—畜牧学]