高浓度葡萄糖对小鼠囊胚细胞线粒体和细胞色素氧化酶的影响

目的:探讨高浓度葡萄糖对小鼠囊胚细胞线粒体和细胞色素氧化酶的影响,探讨高浓度葡萄糖对小鼠囊胚有氧代谢的影响,为糖尿病妊娠导致胎儿畸形的机制和预防提供理论依据。方法:通过促超排卵,获取妊娠3.5天小鼠囊胚,随机分成三组,即对照组(空白)、低糖组(葡萄糖浓度为7.5 mmol/L)和高糖组(葡萄糖浓度为28.0 mmol/L),分别培养在含0、7.5 mmol/L和28.0 mmol/L葡萄糖的M199培养基中,培养24 h后,然后再吸山囊胚。每组随机吸取30个囊胚用罗丹明123荧光染料标记线粒体和组织化学方法显示细胞色素C氧化酶,观察囊胚细胞线粒体和细胞色素C氧化酶(Cytochrome C Oxidase)的变化。结果:①高糖组小鼠囊胚线粒体功能活性明显降低,荧光密度值较低,且线粒体的位置分布发生了明显变化,空白组和低糖组囊胚胞浆中线粒体分布均匀,荧光密度值较高,线粒体仍具有一定的活性。空白组与低糖组囊胚荧光密度值无显著性差异(P>0.05),高糖组与空白组和低糖组相比均存在显著性差异(P<0.05);②细胞色素氧化酶(COX)活性的变化:高糖组小鼠囊胚胞浆淡染,反应产物呈浅黄色,COX活性明显降低;空白组和低糖组囊胚胞浆可见明显的棕褐色物质,COX活性较高,统计学分析,空白组与低糖组囊胚灰度值无显著性差异(P>0.05),高糖组与空白组和低糖组相比均存在显著性差异(P<0.05)。结论:高浓度葡萄糖对小鼠囊胚线粒体的分布、功能以及COX活性均有影响。

Influence of mitochondrial and COX in mouse blastocysts induced by high-glucose

Objective:To investigate the influence of the mitochondria and COX activity in high glucose on mouse blastocysts and to provide more evidence that maternal diabetes impairs embryo development during the earlier phase of gestation.Methods:By superovulated,mouse 3.5 d blastocysts were obtained,expanded blastocysts from different females were pooled and randomly selected for one control group and two experiments groups,which exposed to doses of 0 μmol/L,7.5 μmol/L,and 28.0 μmol/L glucose for 24 h.Blastocysts were stained by Rhodamin 123 to assay the mitochondria respectively.The alteration of COX was examined by histochemistry.Results:①Blastocysts in high glucose showed the decreased activity of mitochondria and the distribution of mitochondria changed,while the mitochondria in blastocysts of control group and low glucose were still active,and the mitochondria were distributed homogeneously in cytoplasm.②The activity of COX decreased in blastocysts of high glucose,which was lower than the blastocysts of control and low glucose group.Conclusion:High glucose can influence the activity of mitochondria and COX,and the distribution of mitochondria.