上海农业学报2017，Vol.33Issue(3)：124-129,6.DOI:10.15955/j.issn1000-3924.2017.03.24
凋亡抑制剂Z-VAD-FMK在猪卵母细胞冷冻保存中的应用
Application of apoptotic inhibitor Z-VAD-FMK in cryopreservation of porcine oocytes
摘要
Abstract
In porcine MII stage,the apoptotic inhibitor Z-VAD-FMK was added to the incubation fluid of porcine oocytes after freezing and thawing,and the effects of Z-VAD-FMK on frozen oocyte were studied by detecting the mitochondrial membrane potential (ΔΨm),early apoptotic status,embryonic development and related gene expression.The results showed:1)Mitochondrial ΔΨm in vitrified group (0.91)was significant lower than that in fresh group (1.33),Z-VAD-FMK addition after warming significantly increased mitochondrial ΔΨm (1.19).2)The early apoptotic rate in vitrified group (57.65％)was significant higher than that in fresh group (8.37 ％),and the survival rate (61.45 ％),cleavage rate (13.18 ％) and blastocyst rate (3.10％) were significant lower than those in fresh group (98.97％,89.41％ and 41.40％).Z-VAD-FMK addition significantly decreased early apoptotic rate (32.82％),and significantly increased survival rate (73.21％),cleavage rate (39.83％) and blastocyst rate (10.79％) respectively.3) Vitrification significantly increased the expression levels of pro-apoptotic related genes and decreased the expression levels of anti-apoptotic genes from both death recepotor and mitochondrial mediated apoptotic pathways.Z-VAD-FMK addition could increase anti-apoptotic gene expression and decrease pro-apoptotic gene expression.In summary,the addition of apoptotic inhibitor Z-VAD-FMK to incubation solution after vitrification could improve mitochondrial ΔΨm,reduce apoptosis and change apoptotic related gene expression levels,and then improve the in vitro developmental competence after parthenogenetic activation.关键词
Z-VAD-FMK/细胞凋亡/玻璃化冷冻/卵母细胞/猪Key words
Z-VAD-FMK/Apoptosis/Vitrification/Oocyte/Porcine分类
农业科技引用本文复制引用
钮莹芳,吴彩凤,张树山,陈亚宁,戴建军,张德福..凋亡抑制剂Z-VAD-FMK在猪卵母细胞冷冻保存中的应用[J].上海农业学报,2017,33(3):124-129,6.基金项目
国家自然科学基金(31372315) （31372315）
国家转基因生物新品种培育科技重大专项(2014ZX08006-005) （2014ZX08006-005）
上海市农业委员会青年人才计划(2014-1-32,2014-1-33) （2014-1-32,2014-1-33）
