p66shc-ROS轴介导猪早期胚胎体外发育阻滞机理的研究

项目名称： p66shc-ROS轴介导猪早期胚胎体外发育阻滞机理的研究

项目编号： No.31301978

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 农业科学

项目作者： 胡军和

作者单位： 湖南人文科技学院

项目金额： 24万元

中文摘要： p66shc是原癌基因SHC编码三个分子量相近的ShcA蛋白质家族中成员之一。研究发现p66shc在由活性氧(reactive oxygen species, ROS)引起细胞凋亡中发挥重要的作用；而在胚胎早期发育过程中，胚胎内ROS是导致胚胎体外发育阻滞的重要原因之一，但是其具体调控机制不详。为此，本研究选择p66shc基因作为引起猪胚胎发育阻滞的靶基因，首先研究发育正常与阻滞的猪胚胎内该基因和ROS水平的关联性，构建p66shc-ROS-胚胎发育的动态模式；然后通过上调（在高氧浓度下等方法）或下调该基因的表达（显微注射RNAi片段，添加抗氧化剂和抗体等）等方法，验证该基因对猪胚胎内ROS水平及发育效率的调控作用。总之，该研究着眼于探索猪胚胎p66shc-ROS-胚胎作用轴调控胚胎发育阻滞的机制，为提高猪胚胎体外发育效率奠定基础。

中文关键词： p66shc；猪；发育阻滞；胚胎发育；活性氧

英文摘要： P66shc (66-kilodahon isoform of Shc gene products) is one of the three ShcA protein family member with similar molecular weight, which is encoded by SHC oncogene of mammal. It has been demonstrated that p66shc protein plays an important role in cell apoptosis caused by reactive oxygen species (ROS); it has also been found that ROS in embryos is one of the most important reasons leading to embryo development block in vitro, but the mechanism is not clear yet. Therefore, this research focuses on p66shc as the research target gene, which is closely related to ROS reaction leading to embryonic development block. In order to prove this hypothesis, the experiments are carried out as following:(1)the levels of ROS and p66shc expression are analyzed between development block and normal development embryos in order to figure out the relation betwwen p66shc gene and ROS level for describing the expression pattern of p66shc-ROS-embryo development;(2)the embryonic ROS level and developing efficiency are researched through microinjecting of p66Shc interference fragment into oocytes or embryos using RNAi technology in vitro; (3)the embryonic ROS level and developing efficiency are researched through adding antioxidants or p66Shc antibody in the culture medium in vitro. In conclusion, the research focus on porcine embryo p66sh

英文关键词： p66shc；porcine；develop block；embryo development；reactive oxygen species