玻璃化冷冻对卵母细胞线粒体损伤及其机理的研究

项目名称： 玻璃化冷冻对卵母细胞线粒体损伤及其机理的研究

项目编号： No.31460644

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 王晓丽

作者单位： 广西大学

项目金额： 50万元

中文摘要： 卵母细胞的冷冻损伤作为卵母细胞冷冻保存的主要障碍越来越受到研究者的关注，目前对卵母细胞冷冻损伤的研究主要集中在微管、微丝和染色体损伤上，而对线粒体损伤的研究报道较少。本课题拟通过多种实验手段检测①冷冻前后卵母细胞内线粒体的结构和分布变化；②测定线粒体膜上的丙二醛、ATP酶及膜电位的变化；③研究冷冻前后卵母细胞内ROS水平的改变、mtDNA损伤情况及ATP含量变化；④探讨ROS对mtDNA复制及损伤修复相关蛋白poly和mtTFA的影响；⑤检测卵母细胞内溶酶体活性变化，以期阐明冷冻是否通过ROS依赖途径调控mtDNA复制及修复相关蛋白的表达及细胞内溶酶体活性变化，导致mtDNA损伤并影响卵母细胞线粒体功能。以澄清卵母细胞玻璃化冷冻后，对线粒体造成的影响，来探讨冷冻对卵母细胞内线粒体功能影响的可能机制，为指导玻璃化冷冻卵母细胞提供新的思路，为进一步探讨影响卵母细胞冷冻保存的机理提供实验参考。

中文关键词： 玻璃化冷冻；卵母细胞；线粒体；损伤

英文摘要： The cryodamage of oocyte was paid more attention as the main obstacle of oocyte cryopreservation. At present, the research of oocyte cryodamage on mitochondrial damage has fewer reports, mainly concentrates on the microtubule, microfilament and chromosome damage. The subject will detect ①The changes of structure and distribution on mitochondria by vitrification; ② The changes of malondialdehyde, ATPase and the membrane potential on mitochondrial membrane; ③The changes of the level of ROS, mtDNA damage and ATP content in the oocyte by vitrification; ④Effect of ROS on poly and mtTFA protein that related to mtDNA replication and injury repair; ⑤The change of lysosome activity in the oocyte, in order to clarify whether refrigeration regulates the expression of mtDNA replication and repair related proteins and the change of lysosome activity in the cell through ROS-depend pathway, thus leading to mtDNA damage and affecting the mitochondrial function of the oocyte. By clarifying the effects on the mitochondria after vitrification of oocyte to discuss the possible mechanism of effect of refrigeration on mitochondrial function in oocyte. It provides new thoughts to guide the vitrification of oocyte and experimental references for further discussion of the mechanism that affect oocyte cryopreservation.

英文关键词： Vitrification;oocyte;mitochondrion;damage