活性氧对鼠早期胚胎抗氧化酶基因表达的影响

发布时间：2018-05-24 19:22

本文选题：小鼠 + 胚胎　；参考：《延边大学》2015年硕士论文

【摘要】：本实验的目的为了检测鼠早期胚胎内过氧化物酶体数量的变化及过氧化物酶体中抗氧化酶基因表达量的变化,并检测了活性氧对鼠胚胎2-细胞时期过氧化物酶体数量的变化及过氧化物酶体中抗氧化酶基因表达量的影响。能够为哺乳动物早期胚胎体外培养抗氧化机制的研究奠定基础。通过抗氧化酶RNA的表达来判断抗氧化酶的表达,并用过氧化物酶体膜蛋白-20来标记过氧化物酶体,从而检测过氧化物酶体数量的变化。通过本实验得到以下分祈结果：1.MnSOD、GPX-1、GPX-4目的基因在鼠胚胎4细胞时期表达量最高,显著高于卵母细胞和1-细胞、2-细胞时期。CAT、GPX-2目的基因在卵母细胞表达量最高,显著高于胚胎1-细胞、2-细胞、4-细胞时期。Cu-Zn SOD目的基因在2-细胞表达量最高,显著高于卵母细胞和1-细胞、4-细胞时期。GPX-3目的基因在卵母细胞、1-细胞、2-细胞、4-细胞时期之间表达无明显差异。2.30、50μmol/L两组的4-细胞率、囊胚率显著低于0、10μmol/L两组,但是,50μmol/L组的卵裂率显著低于其他三组。通过Image软件分析图片得出结果,H202处理组胚胎内ROS水平显著高于体内、体外发育两组,同时体外发育组ROS水平显著高于体内发育。3.H202处理组PMP-20表达量高,Cu-ZnSOD、GPX-3基因表达量高,MnSOD、GPX-2基因表达量低,而GPX-4基因表达量在体外培养组表达量最高,GPX-1基因表达量无显著变化,CAT基因没有表达。结论：不同抗氧化酶基因表达量在鼠早期胚胎发育的不同时期有所不同jMnSOD、 GPX-1、GPX-4基因随着胚胎的发育表达量不断增高,过氧化物酶体数量同时在增加。
[Abstract]:The purpose of this study was to detect the changes of peroxisome number and the expression of antioxidant enzymes in peroxisome in early mouse embryos. The effects of reactive oxygen species (Ros) on the number of peroxisome and the expression of antioxidant enzymes in peroxisome were investigated. It can lay a foundation for the study of antioxidation mechanism of mammalian early embryo culture in vitro. The expression of antioxidant enzyme (RNA) was determined by the expression of antioxidant enzyme. Peroxisome membrane protein (-20) was used to label peroxisome and the quantity of peroxisome was detected. The results obtained from this experiment were as follows: 1. The highest expression of GPX-4 target gene was found in mouse embryonic 4 cells, which was significantly higher than that in oocytes and 1-cell / 2- cell stage. The expression of GPX-2 gene was the highest in oocytes. The expression of Cu-Zn SOD target gene was significantly higher than that of the embryonic 1-cell / 2-cell / 4- cell stage, and the highest expression of Cu-Zn SOD gene was found in the 2-cell. The expression of GPX-3 target gene was significantly higher than that of oocyte and 1-cell / 4- cell stage. There was no significant difference in the expression of the target gene between the oocytes and the oocytes in the oocyte 1-cell and 2-cell / 4- cell phase. The blastocyst rate was significantly lower than that in the 0 ~ 10 渭 mol/L group, and the blastocyst rate was significantly lower in the two groups. However, the cleavage rate of 50 渭 mol/L group was significantly lower than that of the other three groups. The results of Image software analysis showed that the level of ROS in embryos of H202 treatment group was significantly higher than that in vivo. At the same time, the level of ROS in the in vitro development group was significantly higher than that in the in vivo development.3.The PMP-20 expression level in the treatment group was higher than that in the in vivo development.3. However, there was no significant change in the expression of GPX-4 gene in vitro. Conclusion: the expression of different antioxidant enzyme genes is different at different stages of mouse embryonic development. The expression of GPX-1 GPX-4 gene increases with the development of embryo, and the number of peroxisome increases simultaneously.
【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S814

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