单宁酸对猪卵母细胞体外成熟及胚胎发育能力的影响

Effects of Tannin Acid on the in vitro Maturation andEmbryonic Development of Porcine Oocytes

研究旨在探讨单宁酸对猪卵母细胞体外成熟质量及其胚胎发育能力的影响。在猪卵丘卵母细胞复合体(COCs)体外成熟培养液中添加不同浓度(0、1、10、100μg/mL)单宁酸培养42 h后,检测COCs的扩散程度和卵丘细胞扩散指数,统计COCs的体外成熟率,检测成熟卵母细胞内谷胱甘肽(glutathione,GSH)、活性氧(reactive oxygen species,ROS)和生长分化因子9(growth differentiation factor 9,GDF9)的水平,并统计孤雌激活及体外受精胚胎48和168 h的卵裂率、囊胚率及囊胚总细胞数。结果显示,与对照组相比,10μg/mL单宁酸组卵丘细胞扩散指数显著提高(P<0.05),100μg/mL单宁酸组显著降低(P<0.05);1和10μg/mL单宁酸组卵母细胞成熟率差异不显著(P>0.05),100μg/mL单宁酸组卵母细胞成熟率显著降低(P<0.05);1和10μg/mL单宁酸组GSH和GDF9水平显著提高(P<0.05),ROS水平显著降低(P<0.05)。孤雌胚胎和体外受精胚胎发育能力结果显示,与对照组相比,各单宁酸组卵裂率差异不显著(P>0.05),10μg/mL单宁酸组孤雌胚胎囊胚率及体外受精胚胎囊胚率显著提高(P<0.05),100μg/mL单宁酸组孤雌胚胎囊胚细胞数及体外受精胚胎囊胚细胞数均显著低于其他各组(P<0.05)。以上结果表明,10μg/mL单宁酸可通过提高卵丘细胞扩散能力及GSH和GDF9水平、降低卵母细胞内ROS水平,改善猪卵母细胞成熟质量,提高孤雌胚胎及体外受精胚胎的发育能力。

The aim of this study was to investigate the effect of tannin acid on porcine in vitro maturation(IVM)and their subsequent embryonic development.Thus,the porcine cumulus-oocyte complexes were supplemented with different concentration(0,1,10 and 100μg/mL)of tannin acid during 42 h IVM,and then cumulus cells expansion,oocyte maturation rate,and the level of glutathione(GSH),reactive oxygen species(ROS)and growth differentiation factor 9(GDF9)in oocytes were evaluated.Moreover,parthenogenetic activation(PA)and in vitro fertilization(IVF)were carried out,and cleavage rate,blastocyst formation rate and total cell number of blastocyst were evaluated.The results showed that when compared with the control group,10μg/mL tannin acid significantly increased the cumulus expansion index and 100μg/mL tannin acid significantly decreased it(P<0.05).There was no significant difference in oocyte maturation rate between control,1 and 10μg/mL tannin acid groups(P>0.05),while 100μg/mL tannin acid significantly decreased maturation rate(P<0.05).Moreover,the levels of GSH and GDF9 were dramatically enhanced(P<0.05),and the level of ROS was dramatically reduced by 1 and 10μg/mL tannin acid supplementation(P<0.05).The results of development ability of parthenogenetic embryos and in vitro fertilization embryos showed that there was no significant difference in cleavage rate between tannic acid groups and control group(P>0.05).The blastocyst rate of parthenogenetic embryos and in vitro fertilization embryos in 10μg/mL tannic acid group were significantly increased(P<0.05).The number of blastocyst cells in parthenogenetic embryos and in vitro fertilization embryos in 100μg/mL tannic acid group was significantly lower than that in other groups(P<0.05).Taken together,10μg/mL tannin acid improved oocyte quality and the subsequent embryo developmental competence by increasing cumulus cells expansion and the GSH and GDF9 levels,and reducing the ROS level of the porcine oocyte.