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目的:测定葡萄糖培养早期胚胎体外发育各阶段的活性氧(ROS)水平,探讨葡萄糖对小鼠早期胚胎体外发育产生阻滞作用的机制。方法:通过在改良的CZB(m-CZB)培养液中添加不同浓度(分别为0、5、10、15、20、25、30mmol/L)的葡萄糖培养小鼠早期胚胎,根据其体外发育的不同阶段,用分子探针氢化乙啶(hydroethidine,HE)和紫外分光光度法分别检测小鼠胚胎内、外ROS含量。结果:葡萄糖浓度达到15mmol/L时囊胚发育率开始出现明显下降(P<0.05),相同浓度培养的小鼠胚胎在2-细胞晚期阶段胚胎内部超氧阴离子(O2·-)和培养液中过氧化氢(H2O2)含量均明显升高(P<0.05),而发育其他阶段胚胎内部的O2·-与对照组(0mmol/L葡萄糖组)比较均无统计学差异(P>0.05)。结论:高浓度葡萄糖可能在2-细胞晚期阶段诱导ROS增多,致使小鼠早期胚胎体外发育受阻。
OBJECTIVE: To determine the level of reactive oxygen species (ROS) in different stages of early embryos cultured with glucose, and to explore the mechanism of glucose blockade on mouse embryos in vitro. Methods: The early embryos of mice were cultured in modified CZB (m-CZB) medium with different concentrations of glucose (0, 5, 10, 15, 20, 25 and 30 mmol / L, respectively) In different stages, the contents of ROS in mouse embryos were detected by molecular probe hydroethidine (HE) and ultraviolet spectrophotometry respectively. Results: The blastocyst rate of 15 mmol / L glucose began to decline significantly (P <0.05). The same concentration of mouse embryos cultured in the late stage of 2-cell embryo superoxide anion (O2 · -) and culture medium (P <0.05). However, there was no significant difference (P> 0.05) between O2 · - in embryos at other stages of development and control group (0 mmol / L glucose). CONCLUSION: High concentration of glucose may induce ROS increase in the late phase of 2-cell, which hinders the development of mouse early embryos in vitro.