核受体NR4A1介导猪卵巢黄体退化的作用机制研究

项目名称： 核受体NR4A1介导猪卵巢黄体退化的作用机制研究

项目编号： No.31501956

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 畜牧学与草地科学

项目作者： 茆达干

作者单位： 南京农业大学

项目金额： 20万元

中文摘要： 哺乳动物卵巢黄体对母畜正常繁殖周期的调控具有重要作用。PGF2a作为一种主要的溶黄体因子，可诱导黄体发生功能性和结构性退化，但其具体的分子机制还不清楚。核受体NR4A1作为早期反应基因，可被多种生理刺激快速诱导，参与机体的糖脂代谢、类固醇生成、细胞凋亡等调控过程。我们前期的初步研究表明PGF2a处理诱导了猪卵巢黄体中NR4A1蛋白表达的上调，但NR4A1在猪卵巢黄体中的具体作用还不明确。本项目拟通过基因过表达与RNA干涉方法，应用实时定量PCR、WB、IF、染色质免疫沉淀、启动子克隆及活性分析等技术，从在体-细胞-分子水平上研究NR4A1对猪黄体细胞类固醇合成通路和细胞凋亡的影响，揭示NR4A1介导的黄体类固醇生成细胞参与猪卵巢黄体退化的作用机制。通过以上研究，进一步明确PGF2a诱导猪黄体退化的分子机制，深入理解猪性腺功能的调控机制，对提高猪的繁殖力，促进可持续生产具有重要意义。

中文关键词： 卵巢；；黄体；生殖内分泌；细胞凋亡；信号传导

英文摘要： Mammalian corpus luteum plays an important role in the regulation of reproductive cyclicity. Prostaglandin F 2alpha, as a major luteolytic factor, can induce luteal functional and structural regression, but its molecular mechanism is not clear. Nuclear receptor NR4A1, as an early response gene, can be induced by different stimuli, and is involved in glucose and lipid metabolism, steroidogensis, apoptosis, and so on. Our prelimary results have shown that PGF2a treatment induced NR4A1 expression in the porcine luteal cells, but the detailed mechanism in porcine luteolysis is not yet known. In this project, gene overexpression and RNA interference, accompanied with real time-PCR, Western blot, immunoflurorescence, chromatin immunoprecitation, promoter cloning and activity analysis techniques will be applied to study the effect of NR4A1 on luteal steroidogenic signal pathway and cell apoptosis at the In vivo - Cell- Molecular levels, therefore to reveal the mechanism of NR4A1- mediated luteolysis in pigs.These studies will provide a deeper understanding of the molecular mechanism about porcine luteolysis induced by PGF2a and contribute new information to fill a critical gap in our understanding of mechanisms responsible for regulating porcine gonadal function. This knowledge is needed to promote porcine fecundity and sustainable production.

英文关键词： ovary;corpus luteum;reproductive endocrine;cell apoptosis;signal transduction