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葡萄糖在小鼠胚胎2-细胞晚期诱导活性氧增多抑制胚胎体外发育
张雷1,2,李钟淑2,方南洙2*
0
(1.铜仁学院生物化学系动物实验室,铜仁 554300;2.延边大学农学院动物遗传育种与繁殖实验室,龙井133400)
摘要:
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、30 mmol/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15 mmol/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(0 mmol/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。
关键词:  葡萄糖  胚胎发育  活性氧
DOI:10.3724/SP.J.1008.2009.0888
投稿时间:2008-11-05修订日期:2009-03-10
基金项目:国家自然科学基金(30270954).
Glucose-induced ROS inhibits mouse early embryo development at late 2-cell stage in vitro
ZHANG Lei1,2,LI Zhong-shu2,FANG Nan-zhu2*
(1.Laboratory of Animal,Department of Bio-chemistry,Tongren College,Tongren 554300,China;2.Laboratory of Animal Genetic and Breeding,Agriculture College of Yanbian University,Longjing 133400)
Abstract:
Objective:To measure the levels of reactive oxygen species (ROS) at each development stage of early embryos incubated with glucose in vitro,and to probe into the mechanism by which glucose blocks the development of mouse early embryos.Methods: Mouse early embryos were cultured with different concentrations of glucose (0,5,10,15,20,25 and 30 mmol/L) added in modified CZB (m-CZB) medium.ROS levels inside and outside of embryos were examined by molecule probes of hydroethidine and spectrophotometry at different stages of embryos.Results: When glucose concentration reached 15 mmol/L,the blastocyst developmental rates decreased significantly compared with control (0 mmol/L glucose,P<0.05); superoxide anion (O2-·) levels inside embryos and hydrogen peroxide (H2O2 ) levels in the medium with 15 mmol/L glucose were significantly higher (P<0.05) than those in medium with no glucose at late 2-cell stage.However,there was no significant difference in O2-·levels in the embryos exposed to 15 mmol/L glucose and no glucose at other growth stages (P>0.05).Conclusion: High glucose may induce ROS production at late 2-cell stage and block the development of mouse embryos.
Key words:  glucose  embryo development  reactive oxygen species