虾青素对猪卵母细胞体外成熟的影响  

Effects of astaxanthin on maturation of porcine oocytes in vitro

作　　者：许建春 汪浩鑫 自永宏 杨小芬 闫茜 陈梦佳 石德顺[1] 陆凤花[1] XU Jianchun;WANG Haoxin;ZI Yonghong;YANG Xiaofen;YAN Xi;CHEN Mengjia;SHI Deshun;LU Fenghua(State Key Laboratory of Conservation and Utilization of Biological Resources in Subtropical Agriculture,Guangxi University,Nanning 530004,China)

机构地区：[1]广西大学亚热带农业生物资源保护与利用国家重点实验室,广西南宁530004

出　　处：《中国兽医学报》2022年第12期2549-2554,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(3176066);广西壮族自治区自然科学基金资助项目(2020JJD130069)。

摘　　要：本研究旨在探讨虾青素(astaxanthin,Ast)对猪卵母细胞体外成熟及孤雌激活胚胎发育的影响,为进一步优化猪卵母细胞体外成熟培养体系提供参考。首先,将GV期猪卵母细胞培养在添加有不同浓度虾青素(0,5,10,20,40μmol/L)的体外成熟液中,40~44 h后统计第一极体排出率。结果显示与对照组相比,各处理组卵母细胞的第一极体排出率均显著升高(P<0.05),其中10μmol/L处理组卵母细胞的第一极体排出率显著高于5,20,40μmol/L处理组(P<0.05)。随后对MⅡ期卵母细胞进行孤雌激活处理,结果发现10μmol/L和20μmol/L处理组的卵裂率和囊胚发育率显著高于对照组(P<0.05),且10μmol/L处理组囊胚发育率显著高于20μmol/L处理组(P<0.05)。最后,检测虾青素处理后MⅡ期卵母细胞的活性氧水平、丙二醛及谷胱甘肽的含量,以及整个胚胎发育阶段中抗氧化酶相关基因SOD1和GPX4的表达变化,发现GV期卵母细胞经10μmol/L虾青素处理后,孤雌激活囊胚的细胞数极显著升高(P<0.01);而MⅡ期卵母细胞的活性氧水平和丙二醛含量与对照组相比极显著降低(P<0.01),谷胱甘肽含量显著升高(P<0.05)。实时荧光定量PCR(qRT-PCR)结果显示,与对照组相比,GPX4基因在10μmol/L处理组MⅡ期卵母细胞中显著升高(P<0.05)。SOD1基因在10μmol/L处理组中MⅡ期卵母细胞、4-细胞和囊胚期的表达极显著高于对照组(P<0.01)。结果表明,卵母细胞体外成熟液中添加适量浓度的虾青素可有效促进卵母细胞的发育潜能。The aim of this study was to investigate the effect of astaxanthin on the in vitro maturation of porcine oocytes and their subsequent lone female activated embryo development,and to provide a reference for further optimization of the porcine oocyte in vitro maturation culture system.First,porcine oocytes at GV stage were cultured in in vitro maturation solutions supplemented with different concentrations of astaxanthin(0,5,10,20 and 40μmol/L),and the first polar body expulsion rates were counted after 40-44 h.The results showed that the first polar body expulsion rates of oocytes in all treatment groups were significantly higher compared with the control group(P<0.05),in which the first polar body expulsion rates of oocytes in the 10μmol/L treatment group were significantly higher than those in the 5μmol/L treatment group.The first polar body excretion rate of oocytes in the 10μmol/L treatment group was significantly higher than that in the 5,20 and 40μmol/L treatment groups(P<0.05).Subsequently,the MⅡ stage oocytes were treated with solitary activation.The results showed that the rates of oogenesis and blastocyst development were significantly higher in the 10μmol/L and 20μmol/L treatment groups than in the control group(P<0.05),and the rate of blastocyst development was significantly higher in the 10μmol/L treatment group than in the 20μmol/L treatment group(P<0.05).Finally,the reactive oxygen level,malondialdehyde and glutathione content of MⅡ stage oocytes,and the expression changes of antioxidant enzyme-related genes SOD1 and GPX4 throughout the embryonic development stage were examined after astaxanthin treatment,and it was found that the cell number of solitary female-activated blastocysts in GV stage oocytes treated with 10μmol/L astaxanthin was extremely significantly increased(P<0.01);while the cell number of MⅡ stage reactive oxygen levels and malondialdehyde content of MⅡ stage oocytes were highly significantly lower(P<0.01)and glutathione content was significantly higher(P<0.05)comp

关 键 词：虾青素 卵母细胞 体外成熟 孤雌胚胎 

分 类 号：S814.6[农业科学—畜牧学]