没食子酸对黄牛卵母细胞体外成熟的影响

doi:10.16431/j.cnki.1671-7236.2018.08.025

摘要/Abstract

摘要：

试验旨在研究没食子酸（gallic acid，GA）对黄牛卵母细胞体外成熟及早期胚胎发育的影响，进一步优化黄牛卵母细胞体外成熟体系。在卵母细胞体外成熟液（M液）中添加不同浓度没食子酸（0、10、30、50、100 μmol/L），成熟22~24 h后，统计卵丘扩展情况及卵母细胞成熟率；同时，对成熟的卵母细胞进行正常体外受精（IVF），统计早期胚胎的分裂率、囊胚率、囊胚卵裂球数及卵裂球细胞凋亡率。根据试验结果，选择最优浓度，使卵母细胞在含该浓度没食子酸的成熟液中成熟24 h后，检测其细胞内的活性氧水平（ROS）和总谷胱甘肽（TGSH）含量。结果显示，M液中添加30 μmol/L没食子酸组卵丘扩展分值和成熟率显著高于对照组（0 μmol/L）（P<0.05），其他处理组与对照组无显著差异（P>0.05）；成熟的卵母细胞体外受精后进行后续胚胎培养，其中10和30 μmol/L组的分裂率均显著高于对照组和100 μmol/L组（P<0.05），50和100 μmol/L组分裂率较对照组也有所提高，但差异不显著（P>0.05）；早期囊胚率统计发现，与对照组相比，30和100 μmol/L能够显著提高囊胚发育率（P<0.05），10和50 μmol/L浓度组则无显著差异（P>0.05）；与对照组相比，30、100 μmol/L没食子酸均能显著提高IVF胚胎的早期囊胚卵裂球数（P<0.05）；但囊胚卵裂球凋亡率与对照组无显著差异（P>0.05）；对卵母细胞内活性氧和总谷胱甘肽含量检测时，发现30 μmol/L没食子酸可显著降低细胞内活性氧水平（P<0.05），且显著提高总谷胱甘肽含量（P<0.05）。综上所述，在黄牛卵母细胞体外成熟液中添加适量的没食子酸能有效降低卵母细胞内活性氧水平，提高总谷胱甘肽含量，进而提高卵母细胞成熟的质量及其后续IVF胚胎发育能力。

关键词: 没食子酸; 体外受精; 黄牛卵母细胞

Abstract:

The aim of this study was to investigate the effects of adding various concentrations of gallic acid (GA) to the maturation medium on the maturation of bovine oocytes and early embryo development in vitro in order to further optimize culture system of bovine oocytes in vitro maturation.Oocytes were matured in vitro on culture medium with different concentrations (0,10,30,50,100 μmol/L) of GA in incubator for 22 to 24 h.Subsequently,the maturation rate of oocyte and the expansion of cumulus cell were calculated,and the cleavage rates,the blastocyst rates,the number of blastomere and the apoptosis rate of blastomere were evaluated after in vitro fertilization (IVF).According to the above test results,oocytes were matured in vitro on culture medium with the optimal concentration of GA for 24 h,and then the intracellular reactive oxygen species (ROS) level and total glutathione level were detected.The results showed that the mature medium with 30 μmol/L GA could significantly increase the oocytes maturation rates and the sore of expansion of cumulus cell than those of the control group (P<0.05),but there was no significant difference between other treatment groups and control group (P>0.05).The cleavage rates of 10 and 30 μmol/L GA groups were significantly increased than 0 and 100 μmol/L GA groups(P<0.05),while there were no significant differences among 0,50,100 μmol/L GA groups (P>0.05).The blastocyst rate of 30 and 100 μmol/L GA groups were significantly improved than that of control group (P<0.05),but 10 and 50 μmol/L GA groups had no significant differences with control group (P>0.05).The No.of blastomere in 30 and 100 μmol/L GA groups were significantly higher than that of the control group (P<0.05),however,the apoptosis rate of blastomere in each group had no significant difference (P>0.05).The detection of the ROS level and total glutathione content in oocyte showed that the levels of ROS were significantly decreased and the content of total glutathione increased significantly in 30 μmol/L GA group (P<0.05).These results suggested that adding GA during IVF might reduce oxidative stress by decreasing ROS levels directly and increasing total glutathione levels in oocytes,thus improve the quality of oocyte maturation and its subsequent IVF embryos developmental ability.

Key words: gallic acid (GA); in vitro fertilization (IVF); bovine oocytes

中图分类号:

S814

黄永军, 李小凤, 文冬梅, 佘春, 邓凯, 潘尔科, 邵齐明, 石德顺, 韦英明, 陆凤花. 没食子酸对黄牛卵母细胞体外成熟的影响[J]. 《中国畜牧兽医》, 2018, 45(8): 2247-2254.

HUANG Yongjun, LI Xiaofeng, WEN Dongmei, SHE Chun, DENG Kai, PANG Erke, SHAO Qiming, SHI Deshun, WEI Yingming, LU Fenghua. Effects of Gallic Acid on in vitro Maturation of Bovine Oocytes[J]. , 2018, 45(8): 2247-2254.

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