本文選題：小鼠 + 胚胎��； 參考：《延邊大學(xué)》2015年碩士論文

【摘要】：本實驗的目的為了檢測鼠早期胚胎內(nèi)過氧化物酶體數(shù)量的變化及過氧化物酶體中抗氧化酶基因表達量的變化,并檢測了活性氧對鼠胚胎2-細胞時期過氧化物酶體數(shù)量的變化及過氧化物酶體中抗氧化酶基因表達量的影響。能夠為哺乳動物早期胚胎體外培養(yǎng)抗氧化機制的研究奠定基礎(chǔ)。通過抗氧化酶RNA的表達來判斷抗氧化酶的表達,并用過氧化物酶體膜蛋白-20來標(biāo)記過氧化物酶體,從而檢測過氧化物酶體數(shù)量的變化。通過本實驗得到以下分祈結(jié)果：1.MnSOD、GPX-1、GPX-4目的基因在鼠胚胎4細胞時期表達量最高,顯著高于卵母細胞和1-細胞、2-細胞時期。CAT、GPX-2目的基因在卵母細胞表達量最高,顯著高于胚胎1-細胞、2-細胞、4-細胞時期。Cu-Zn SOD目的基因在2-細胞表達量最高,顯著高于卵母細胞和1-細胞、4-細胞時期。GPX-3目的基因在卵母細胞、1-細胞、2-細胞、4-細胞時期之間表達無明顯差異。2.30、50μmol/L兩組的4-細胞率、囊胚率顯著低于0、10μmol/L兩組,但是,50μmol/L組的卵裂率顯著低于其他三組。通過Image軟件分析圖片得出結(jié)果,H202處理組胚胎內(nèi)ROS水平顯著高于體內(nèi)、體外發(fā)育兩組,同時體外發(fā)育組ROS水平顯著高于體內(nèi)發(fā)育。3.H202處理組PMP-20表達量高,Cu-ZnSOD、GPX-3基因表達量高,MnSOD、GPX-2基因表達量低,而GPX-4基因表達量在體外培養(yǎng)組表達量最高,GPX-1基因表達量無顯著變化,CAT基因沒有表達。結(jié)論：不同抗氧化酶基因表達量在鼠早期胚胎發(fā)育的不同時期有所不同jMnSOD、 GPX-1、GPX-4基因隨著胚胎的發(fā)育表達量不斷增高,過氧化物酶體數(shù)量同時在增加。
[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.
【學(xué)位授予單位】：延邊大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S814

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本文編號：1930253