熊果苷对小鼠卵母细胞体外成熟及早期胚胎发育潜能的影响机制

Mechanism of Effect of Arbutin on Mouse Oocyte Maturation and Early Embryonic Development Potential in Vitro

目的探究熊果苷对小鼠卵母细胞体外成熟及早期胚胎发育的影响。方法使用含有不同剂量的熊果苷(0、10、20、40µmol/L,分别设为对照组、10µM组、20µM组、40µM组)培养基对生发泡(germinal vesicle,GV)期小鼠的卵母细胞进行16 h体外培养,监测卵母细胞成熟率和活性氧物质(reactive oxygenspecies,ROS)含量。体外受精后将受精卵转移到相同的培养基中,观察卵裂率和囊胚率的变化。结果补充熊果苷培养基的卵母细胞的体外卵母细胞成熟度高于对照卵母细胞,其中40µM组(100%)明显高于对照组(50%),差异有统计学意义(χ^(2)=10.526,P=0.001)。4组的第一极体排出率比较,差异无统计学意义(P均>0.05)。10µM组、20µM组、40µM组卵母细胞内的ROS荧光强度均低于对照组,差异有统计学意义(P均<0.05)。4组囊胚率比较,差异无统计学意义(P均>0.05)。结论在成熟液中添加适量熊果苷能降低卵母细胞中的ROS水平,但对小鼠胚胎发育无明显影响。

Objective To explore the effect of arbutin on mouse oocyte maturation and early embryonic development in vitro.Methods Oocytes from mice at the germinal vesicle(GV)stage were cultured in vitro for 16 h using medium containing different doses of arbutin(0,10,20,and 40μmol/L,set up as control,10μM,20μM,and 40μM groups,respectively).Oocyte maturation rate and reactive oxygen species(ROS)levels were monitored.After in vitro fertilization,fertilized eggs were transferred to the same medium and changes in cleavage rate and blastocyst rate were observed.Results There was no statistically significant difference in the number of oocytes among the four groups(all P>0.05).The in vitro oocyte maturation of oocytes supplemented with arbutin medium was higher than that of control oocytes,and the 40μM group(100%)was significantly higher than the control group(50%),the difference was statistically significant(χ^(2)=10.526,P=0.001).There was no statistically significant difference in the first polar body discharge rate among the 4 groups(all P>0.05).The ROS fluorescence intensity in oocytes of 10μM group,20μM group and 40μM group was lower than that of the control group,and the difference was statistically significant(all P<0.05).Conclusion The addition of arbutin in maturation fluid plays an important role in reducing intracellular stress,protecting cells from apoptosis and increasing the quality of egg cell maturation,and can reduce the ROS level in oocytes,but has no significant effect on mouse embryo development.