摘要
将收集的猪COCs置于添加不同浓度(0.0,0.3,0.6,1.2μmol/L)甘草酸单铵盐(monoammonium glycyrrhizinate,MAG)的卵母细胞体外成熟培养液中培养46 h,统计成熟率,通过免疫荧光染色检测成熟卵母细胞ROS表达水平;对成熟卵母细胞体外受精,于胚胎培养液内体外培养48,120 h,分别统计体外受精胚胎卵裂率、囊胚率,并用Hochest荧光染色检测囊胚总细胞数;结合成熟率及IVF胚胎发育囊胚率选定最佳添加浓度,在体外成熟培养液中添加最适浓度MAG,以0μmol/L MAG为对照组,体外培养46 h后,利用免疫荧光染色检测猪成熟卵母细胞的线粒体膜电位水平和细胞凋亡水平。结果显示,与对照组相比,不同浓度MAG处理组猪卵母细胞体外成熟率均有所提高,但差异均不显著(P>0.05);不同浓度MAG添加均可降低猪卵母细胞ROS水平,与对照组相比,0.3,0.6μmol/L组差异显著(P<0.05),1.2μmol/L组差异极显著(P<0.01)。0.3μmol/L添加组显著提高了猪体外受精囊胚率(P<0.05),但不同MAG添加组对胚胎卵裂率、囊胚总细胞数均无显著影响(P>0.05)。与对照组相比,0.3μmol/L MAG添加组卵母细胞线粒体膜电位水平显著升高(P<0.05),细胞凋亡水平显著降低(P<0.05)。结果表明,0.3μmol/L MAG可提高猪卵母细胞线粒体活性、降低其氧化应激水平和凋亡水平,提高卵母细胞体外成熟质量,从而提高猪体外受精胚胎发育能力。
The collected porcine COCs were placed in the in vitro maturation medium of porcine oocytes supplemented with different concentrations(0.0,0.3,0.6,1.2 μmol/L) of monoammonium glycyrrhizinate(MAG) for 46 h,the maturity rate was calculated,and the expression level of ROS in mature oocytes was detected by immunofluorescence staining.In vitro fertilization of mature oocytes,cleavage rate and blastocyst rate of IVF embryos were measured at 48 h and 120 h,respectively.Hochest fluorescence staining was used to detect the total number of blastocyst cells.The optimal concentration of MAG was added into the in vitro maturation medium,and 0.0 μmol/L MAG was used as the control group.After 46 h in vitro culture,the mitochondrial membrane potential and apoptosis of porcine mature oocytes were detected by immunofluorescence staining.Compared with the control group,the in vitro maturation rates of porcine oocytes in different MAG treatment groups were increased,but the differences were not significant(P>0.05).The levels of ROS in porcine oocytes were decreased by MAG supplementation at different concentrations,and there were significant differences in 0.3,0.6μmol/L groups(P<0.05).Compared with the control group,the mitochondrial membrane potential level of oocytes in 0.3μmol/L MAG supplemental group was significantly increased(P<0.05),and the apoptosis level was significantly decreased(P<0.05).0.3μmol/L MAG could increase mitochondrial activity,reduce the oxidative stress and apoptosis level of porcine oocytes,improve the quality of in vitro maturation of porcine oocytes,and significantly improve the developmental ability of IVF embryos.
作者
张碧菡
赵宝宝
高静
沈成龙
王勇胜
马会明
卿素珠
ZHANG Bihan;ZHAO Baobao;GAO Jing;SHENG Chenglong;WANG Yongsheng;MA Huiming;QING Suzhu(College of Veterinary Medicine,Northwest A&F University,Shaanxi 712100,China;Ministry of Education Key Laboratory for Fertility Preservation and Maintenance,Ningxia Medical University,Yinchuan 750000,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2023年第11期2361-2367,共7页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(K3010322111)。
关键词
甘草酸单铵盐
猪卵母细胞
体外成熟
体外受精
胚胎发育
monoammonium glycyrrhizinate
porcine oocyte
in vitro maturantion
in vitro fertiliza-tion
embryonic development
