本文選題：孤雌胚胎 + 抗氧化劑��； 參考：《華北理工大學(xué)》2017年碩士論文

【摘要】：目的研究不同濃度谷胱甘肽(GSH)、維生素E(VE)、維生素C(VC)對(duì)小鼠孤雌激活胚胎體外發(fā)育的影響,探討三種抗氧化劑提高孤雌胚胎發(fā)育能力的適宜濃度,為改善胚胎體外培養(yǎng)體系及提高孤雌胚胎體外發(fā)育潛能提供實(shí)驗(yàn)依據(jù)。方法本實(shí)驗(yàn)以CD-1?小鼠為研究對(duì)象,根據(jù)不同的抗氧化劑分為三個(gè)實(shí)驗(yàn)組,每組設(shè)置4個(gè)濃度點(diǎn),每個(gè)濃度點(diǎn)隨機(jī)分配10只雌性小鼠。谷胱甘肽組設(shè)置0mmol/L(空白對(duì)照組)、0.5mmol/L、1mmol/L、1.5mmol/L四個(gè)濃度點(diǎn);維生素E組設(shè)置0μmol/L(空白對(duì)照組)、50μmol/L、100μmol/L、150μmol/L四個(gè)濃度點(diǎn);維生素C組設(shè)置0μmol/L(空白對(duì)照組)、100μmol/L、200μmol/L、300μmol/L四個(gè)濃度點(diǎn)。對(duì)小鼠促排卵后取卵團(tuán),體外進(jìn)行孤雌激活。將激活后的卵母細(xì)胞分配到含有不同濃度抗氧化劑培養(yǎng)液中,觀察、計(jì)數(shù)各發(fā)育階段卵裂率、囊胚形成率、囊胚孵出率等指標(biāo),并在卵裂期和囊胚期階段抽取樣本在透射電鏡下觀察抗氧化劑對(duì)孤雌胚胎細(xì)胞微結(jié)構(gòu)的影響。結(jié)果1谷胱甘肽(GSH)組中,各濃度點(diǎn)小鼠孤雌胚胎卵裂率比較差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),囊胚形成率和囊胚孵出率之間差異均有統(tǒng)計(jì)學(xué)意義(P0.05);GSH濃度為1mmol/L時(shí)所獲得的卵裂率、囊胚形成率、囊胚孵出率較高。透射電鏡下觀察添加谷胱甘肽實(shí)驗(yàn)組比對(duì)照組細(xì)胞器結(jié)構(gòu)良好,線粒體空泡及細(xì)胞凋亡現(xiàn)象較少。2維生素E(VE)組中,各濃度點(diǎn)卵裂率、囊胚形成率、囊胚孵率比較差異均有統(tǒng)計(jì)學(xué)意義(P0.05);VE濃度為100μmol/L獲得的卵裂率、囊胚形成率、囊胚孵出率較高。透射電鏡下觀察添加VE實(shí)驗(yàn)組與對(duì)照組比較細(xì)胞微結(jié)構(gòu)良好,線粒體空泡及細(xì)胞凋亡現(xiàn)象少。3維生素C(VC)組中,小鼠孤雌胚胎各濃度點(diǎn)卵裂率、囊胚形成率、囊胚孵出率比較差異均有統(tǒng)計(jì)學(xué)意義(P0.05);200μmol/L組的卵裂率、囊胚形成率、囊胚孵出率較高,300μmol/L組的卵裂率、囊胚形成率、囊胚孵出率較低。透射電鏡下觀察200μmol/L組與對(duì)照組比較細(xì)胞器結(jié)構(gòu)良好,300μmol/L組比對(duì)照組線粒體空泡及細(xì)胞凋亡現(xiàn)象增多。4對(duì)1.0mmol/L谷胱甘肽、100μmol/L維生素E、200μmol/L維生素C進(jìn)行比較,200μmol/L VC獲得的卵裂率、囊胚形成率和囊胚孵出率均最高,與其它兩種濃度的抗氧化劑比較差異有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論1胚胎培養(yǎng)液中添加一定濃度的抗氧化劑對(duì)小鼠孤雌胚胎體外發(fā)育有促進(jìn)作用,1mmol/L GSH、100μmol/L VE、200μmol/L VC是提高發(fā)育潛能的適宜濃度。2300μmol/L VC顯著降低小鼠孤雌胚胎的發(fā)育潛能,提示高濃度的VC可能對(duì)孤雌胚胎產(chǎn)生不良影響。3三種不同濃度抗氧化劑中,200μmol/L的VC對(duì)小鼠孤雌胚胎體外發(fā)育促進(jìn)作用最明顯,能夠獲得較高的卵裂率、囊胚形成率及囊胚孵出率。
[Abstract]:Objective to study the effects of different concentrations of glutathione (GSH), vitamin E (VEV) and vitamin C (VC) on the development of mouse parthenogenetic embryos in vitro, and to explore the appropriate concentrations of three antioxidants to improve the ability of parthenogenetic embryos. In order to improve the system of embryo culture in vitro and improve the development potential of parthenogenetic embryos in vitro. Methods CD-1? Mice were divided into three experimental groups according to different antioxidants. Each group was divided into 4 concentration points and 10 female mice were randomly assigned at each concentration point. The glutathione group had four concentration points of 0 mmol / L (0.5 mmol / L / L), 1.5 mmol / L of 1 mmol / L, 0 渭 mol / L of vitamin E (50 渭 mol / L of 100 渭 mol / L 路L / L) and 150 渭 mol / L / L of 100 渭 mol / L in the control group, and 0 渭 mol / L / L of 100 渭 mol / L (100 渭 mol / L, 200 渭 mol / L, 300 渭 mol / L) of 100 渭 mol / L, 100 渭 mol / L, 300 渭 mol / L, respectively. Parthenogenetic activation was performed in vitro after ovulation stimulation. The activated oocytes were distributed into the medium containing different concentrations of antioxidants. The cleavage rate, blastocyst formation rate, blastocyst hatching rate and so on were observed and counted. The effects of antioxidants on the microstructures of parthenogenetic embryonic cells were observed under transmission electron microscope. Results 1There was no significant difference in cleavage rate of parthenogenetic embryos in glutathione (GSH) group. There were significant differences between blastocyst formation rate and blastocyst hatching rate. The cleavage rate and blastocyst formation rate were obtained when the concentration of GSH was 1 mmol / L. The hatching rate of blastocysts is higher. Under transmission electron microscope (TEM), the cleavage rate and blastocyst formation rate were observed at different concentration points in the experimental group with glutathione as compared with the control group. The difference of blastocyst hatching rate was statistically significant. The cleavage rate, blastocyst formation rate and blastocyst hatching rate of 100 渭 mol / L blastocyst were higher. Transmission electron microscope (TEM) was used to observe the cleavage rate and blastocyst formation rate of mouse parthenogenetic embryos at different concentration points in comparison with control group. There were significant differences in blastocyst hatching rate, blastocyst formation rate, blastocyst hatching rate, blastocyst formation rate, blastocyst hatching rate and blastocyst hatching rate. The cleavage rate of 200 渭 mol / L vitamin E 200 渭 mol / L vitamin C in the 200 渭 mol / L group was higher than that in the control group (200 渭 mol / L VC) by transmission electron microscope. The number of mitochondria vacuoles and cell apoptosis in the 300 渭 mol / L group was higher than that in the control group, and the percentage of 100 渭 mol / L glutathione 100 渭 mol / L vitamin C was higher than that in the control group. The blastocyst formation rate and blastocyst hatching rate were the highest, and the difference was statistically significant compared with the other two concentrations of antioxidants. Conclusion (1) adding a certain concentration of antioxidants in embryonic culture medium can promote the development of mouse parthenogenetic embryos in vitro, and 1 mmol / L GSHH 100 渭 mol / L VEX 200 渭 mol / L VC is a suitable concentration to increase the developmental potential. 2300 渭 mol / L VC can significantly reduce the developmental potential of parthenogenetic embryos. It is suggested that high concentration of VC may have adverse effects on parthenogenetic embryos. 3 among the three different concentrations of antioxidants, 200 渭 mol / L VC can promote the development of mouse parthenogenetic embryos in vitro, and can obtain higher cleavage rate, blastocyst formation rate and blastocyst hatching rate.
【學(xué)位授予單位】：華北理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R714.8

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