自噬在小鼠卵母细胞老化中作用的初步研究

发布时间：2018-04-11 18:38

本文选题：卵母细胞 + 老化　；参考：《山东农业大学》2015年硕士论文

【摘要】：卵母细胞老化过程中氧自由基累积导致线粒体、内质网等细胞器功能受损,细胞内Ca2+平衡失调,MPF活性下降,卵母细胞的激活敏感性升高,胚胎发育能力下降。自噬是吞噬自身细胞质蛋白或细胞器并使其包被进入囊泡,与溶酶体融合形成自噬溶酶体,降解其所包裹的内容物的过程,藉此实现细胞本身的代谢需要和某些细胞器的更新。研究发现自噬是胚胎发育所必需的,并且在卵母细胞体外成熟期间激活自噬能够提高胚胎发育能力,但卵母细胞老化过程中自噬的变化及作用研究较少。因此我们以小鼠为模型,研究体内成熟卵母细胞在体内和体外老化过程中自噬的变化规律,以及自噬对卵母细胞老化和发育的影响。结果如下：1、小鼠卵母细胞体内老化12h后自噬降低,老化24h后自噬不再下降。2、小鼠卵母细胞体外老化12h期间在H202条件培养基中培养,与对照组相比自噬下降；添加MG132组自噬则高于对照组。3、小鼠卵母细胞体外老化12h期间,添加自噬激活剂Rapamycin,自噬增加,激活率下降,胚胎发育能力升高；而添加自噬抑制剂3-MA,自噬下降,激活率增加,胚胎发育能力降低。4、小鼠卵母细胞体外老化12h后自噬降低,老化24h后自噬不再下降。5、小鼠卵母细胞体外老化0h时或老化24h时添加3-MA,降低卵母细胞碎裂率；而在卵母细胞老化12h或24h期间添加3-MA,不影响卵母细胞碎裂。6、小鼠卵母细胞体外老化期间,添加Rapamycin,不影响卵母细胞碎裂。总之,卵母细胞无论在体内还是在体外老化过程中自噬下降,老化24h后自噬不再下降。卵母细胞体外老化12h期间,激活自噬,降低激活率,提高发育能力；抑制自噬,增加激活率,降低发育能力。自噬不影响卵母细胞碎裂。
[Abstract]:During oocyte aging, the accumulation of oxygen free radicals resulted in damage of mitochondria, endoplasmic reticulum and other organelles, decreased the activity of Ca2, increased the sensitivity of oocytes to activation, and decreased the ability of embryonic development.Autophagy is the process of phagocytosis of its own cytoplasmic protein or organelle and its encapsulation into vesicles, fusion with lysosome to form autophagy lysosomes, and degradation of the contents contained therein, thereby realizing the metabolic needs of the cells themselves and the renewal of some organelles.It was found that autophagy is necessary for embryonic development, and activation of autophagy during oocyte maturation in vitro can improve the embryonic development ability, but there is little research on the changes and effects of autophagy during oocyte aging.So we used mouse model to study the changes of autophagy during in vivo and in vitro aging and the effect of autophagy on the aging and development of oocytes.The results were as follows: (1) the autophagy of mouse oocytes decreased after 12h aging and stopped decreasing after 24h aging. The mouse oocytes were cultured in H202 medium during 12h aging in vitro, and the autophagy decreased compared with the control group.During 12h aging of mouse oocytes, the rate of autophagy increased, the rate of autophagy decreased and the ability of embryonic development increased, but the rate of autophagy decreased and the activation rate increased.The development ability of mouse oocytes decreased, the autophagy of mouse oocytes decreased after aging in vitro for 12 h, and the autophagy did not decrease at 24 h after aging. The rate of oocyte fragmentation was reduced by adding 3-MAat at 0 h or 24 h after aging in vitro.The addition of 3-MAduring 12h or 24h of oocyte aging did not affect the cleavage of oocytes. However, during the aging of mouse oocytes in vitro, Rapamycin was added and oocyte fragmentation was not affected.In short, the autophagy of oocytes decreased during in vivo and in vitro aging, and no longer decreased after 24 h aging.During 12h aging in vitro, oocytes activated autophagy, decreased activation rate and increased developmental ability, inhibited autophagy, increased activation rate and decreased developmental ability.Autophagy does not affect oocyte fragmentation.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S814

【参考文献】