抗氧化剂对小鼠孤雌胚胎体外发育的影响

发布时间：2018-06-13 02:31

本文选题：孤雌胚胎 + 抗氧化剂　；参考：《华北理工大学》2017年硕士论文

【摘要】：目的研究不同浓度谷胱甘肽(GSH)、维生素E(VE)、维生素C(VC)对小鼠孤雌激活胚胎体外发育的影响,探讨三种抗氧化剂提高孤雌胚胎发育能力的适宜浓度,为改善胚胎体外培养体系及提高孤雌胚胎体外发育潜能提供实验依据。方法本实验以CD-1?小鼠为研究对象,根据不同的抗氧化剂分为三个实验组,每组设置4个浓度点,每个浓度点随机分配10只雌性小鼠。谷胱甘肽组设置0mmol/L(空白对照组)、0.5mmol/L、1mmol/L、1.5mmol/L四个浓度点;维生素E组设置0μmol/L(空白对照组)、50μmol/L、100μmol/L、150μmol/L四个浓度点;维生素C组设置0μmol/L(空白对照组)、100μmol/L、200μmol/L、300μmol/L四个浓度点。对小鼠促排卵后取卵团,体外进行孤雌激活。将激活后的卵母细胞分配到含有不同浓度抗氧化剂培养液中,观察、计数各发育阶段卵裂率、囊胚形成率、囊胚孵出率等指标,并在卵裂期和囊胚期阶段抽取样本在透射电镜下观察抗氧化剂对孤雌胚胎细胞微结构的影响。结果1谷胱甘肽(GSH)组中,各浓度点小鼠孤雌胚胎卵裂率比较差异无统计学意义(P0.05),囊胚形成率和囊胚孵出率之间差异均有统计学意义(P0.05);GSH浓度为1mmol/L时所获得的卵裂率、囊胚形成率、囊胚孵出率较高。透射电镜下观察添加谷胱甘肽实验组比对照组细胞器结构良好,线粒体空泡及细胞凋亡现象较少。2维生素E(VE)组中,各浓度点卵裂率、囊胚形成率、囊胚孵率比较差异均有统计学意义(P0.05);VE浓度为100μmol/L获得的卵裂率、囊胚形成率、囊胚孵出率较高。透射电镜下观察添加VE实验组与对照组比较细胞微结构良好,线粒体空泡及细胞凋亡现象少。3维生素C(VC)组中,小鼠孤雌胚胎各浓度点卵裂率、囊胚形成率、囊胚孵出率比较差异均有统计学意义(P0.05);200μmol/L组的卵裂率、囊胚形成率、囊胚孵出率较高,300μmol/L组的卵裂率、囊胚形成率、囊胚孵出率较低。透射电镜下观察200μmol/L组与对照组比较细胞器结构良好,300μmol/L组比对照组线粒体空泡及细胞凋亡现象增多。4对1.0mmol/L谷胱甘肽、100μmol/L维生素E、200μmol/L维生素C进行比较,200μmol/L VC获得的卵裂率、囊胚形成率和囊胚孵出率均最高,与其它两种浓度的抗氧化剂比较差异有统计学意义(P0.05)。结论1胚胎培养液中添加一定浓度的抗氧化剂对小鼠孤雌胚胎体外发育有促进作用,1mmol/L GSH、100μmol/L VE、200μmol/L VC是提高发育潜能的适宜浓度。2300μmol/L VC显著降低小鼠孤雌胚胎的发育潜能,提示高浓度的VC可能对孤雌胚胎产生不良影响。3三种不同浓度抗氧化剂中,200μmol/L的VC对小鼠孤雌胚胎体外发育促进作用最明显,能够获得较高的卵裂率、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.
【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R714.8

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