组织蛋白酶抑制剂E-64对猪卵母细胞体外发育能力的影响

Effect of E-64 on Developmental Capacity of Porcine Oocytes in vitro

为探明外源性组织蛋白酶抑制剂(E-64)对猪卵母细胞体外发育能力的影响,通过向成熟液中添加1、5、10μmol/L的外源性组织蛋白酶抑制剂,对猪卵丘-卵母细胞复合体(COCs)进行体外成熟培养46 h,统计第一极体率,随后分别进行体外受精和孤雌激活,统计第2天的卵裂率和第7天的囊胚率。结果表明,成熟液中加入1μmol/L E-64的第一极体排出率显著高于未添加的对照组以及5、10μmol/L E-64的添加组。将体外成熟的卵母细胞进行体外受精及孤雌激活,体外受精后胚胎的卵裂率与囊胚率结果均为成熟液中加入1μmol/L E-64的添加组显著高于对照组以及5、10μmol/L E-64的添加组;孤雌激活后,4组间胚胎的卵裂率差异不显著,囊胚率为10μmol/L E-64的添加组显著低于其他3组。因此1μmol/L E-64能促进猪卵母细胞的核成熟,并能进一步提高其体外受精后的卵裂率和体外培养的囊胚率。

The aim of this study was to ascertain the effect of exogenous cathepsin inhibitor （E-64） on the viability of porcine oocytes developed in vitro. The maturation medium was added with 1, 5, 10 μmol/L exogenous cathepsin inhibitors （E-64） respectively, and the porcine cumulus oocyte complexes （COCs） were matured in vitro for 46 hours, the first polar body rate was calculated. Then in vitro fertilization and parthenogenetic activation were conducted, and the cleavage rate and the blastocyst rate were analyzed. The results showed that the first polar body rate, as well as the associated cleavage rate and blastocyst rate in maturation medium added with 1 μmol/L E-64 were significantly higher than that of other three groups. After parthenogenetic activation, there was no significant difference in the cleavage rate among the four groups, while the blastocyst rate in maturation medium added with 10 μmol/L E-64 was significantly lower than that of other three groups. It could be concluded that the suitable supplement of E-64 for in vitro nuclear maturation of porcine oocytes, as well as the cleavage and embryonic development, should be 1 μmol/L.