日粮硒缺乏致雏鸡胰腺萎缩差异基因表达分析及分子网络构建

项目名称： 日粮硒缺乏致雏鸡胰腺萎缩差异基因表达分析及分子网络构建

项目编号： No.31272468

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 赵华

作者单位： 四川农业大学

项目金额： 86万元

中文摘要： 硒在胰腺正常功能维持方面发挥重要作用，雏鸡胰腺是对硒缺乏最敏感的靶器官，缺硒可致其胰腺萎缩，但生物学机制尚不清楚。以鸡为实验动物，采用现代生物学技术探索缺硒致胰腺萎缩的分子生物学机制，具有重要意义。本项目拟采用四川严重缺硒地区日粮，构建缺硒致雏鸡胰腺萎缩模型并与正常硒胰腺进行比较；采用基因芯片技术比较缺硒对雏鸡胰腺基因表达的影响；通过不同层次分子网络的构建对差异表达基因进行分析，确定在调控中起关键作用的基因；采用实时荧光定量PCR技术研究缺硒对胰腺25个硒蛋白基因表达的影响；对筛选出的部分关键基因，制备抗体进行Western-blot分析，考察雏鸡胰腺中相关蛋白表达变化；在此基础上，综合分析差异基因与胰腺功能维持的关系。我国是一个严重贫硒的大国，研究结果可为人和动物的硒营养应用提供理论参考，同时也可望为当今困扰人类健康的与胰腺相关重大疾病（如糖尿病）发生的分子机理研究和预防提供靶标基因。

中文关键词： 硒缺乏；肉鸡；胰腺萎缩；基因芯片；定量PCR

英文摘要： Selenium (Se) plays important roles on maintaining the normal function of pancreas, chicken pancreas is the organ which most sensitive to Se-deficient and dietary Se-deficiency could induce pancreatic atrophy, however, the biological mechanism is unclear. Taking broiler as research materials and applying on advanced biology technology to reveal the biological mechanism associated with the pancreatic atrophy induced by dietary Se-deficiency is of great meaningful. In this project corn and soybean produced in severe Se-deficient area of Sichuan will be take as basal diet (Se-deficiency diet) to develop the chicken pancreatic atrophy model and following experiments will be conducted: 1) Compare gene express profiles of whole-genome for pancreas between the chickens fed on Se-deficiency diet(-Se) and normal Se diet (+Se) by adoption microarray technology; 2) Screen and data mined of the differentially expressed genes through different level molecular network reconstruction and find out the target genes associated with the pancreatic atrophy induced by Se-deficiency;3) Compare the 25 selenoprotein gene express profiles for pancreas between the two group chickens (-Se and +Se) by adoption the Real-time RT-PCR technique;4) Prepared polyclone antibodies for some screened target genes and investigate the effect of dietar

英文关键词： Selenium-deficiency；Chicken；Pancreatic Atrophy；DNA microarray；qPCR