硒蛋白W对鸡缺硒性骨骼肌损伤中钙稳态的调控

项目名称： 硒蛋白W对鸡缺硒性骨骼肌损伤中钙稳态的调控

项目编号： No.31272626

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： 徐世文

作者单位： 东北农业大学

项目金额： 77万元

中文摘要： 鸡缺硒性骨骼肌损伤中存在硒蛋白W含量降低与钙稳态紊乱，硒蛋白W二级结构中存在钙调结构域，但其在钙稳态失衡中的作用机制尚不清楚。本试验首先建立硒蛋白W基因敲除鸡模型和骨骼肌细胞体外培养的硒蛋白W基因敲除和过表达模型，在人工复制鸡缺硒性骨骼肌损伤病例模型基础上，应用RNA干扰和过表达技术、流式细胞术、蛋白印记试验、实时荧光定量PCR检测技术、激光共聚焦显微技术等，通过对骨骼肌内钙稳态、钙敏感性受体含量及其基因mRNA表达、肌浆网钙摄取与释放、钙调蛋白、钙网蛋白、肌浆网相关应激因子（GRP78/Bip、CHOP/GADDl53、XBP-1）、细胞凋亡等的检测，阐明硒蛋白W在鸡缺硒性骨骼肌损伤钙稳态失衡中的作用机制，本项目的实施将有助于加深对该病发病机理的认识，同时也为比较医学在该领域的研究提供新的思路。

中文关键词： 鸡；硒蛋白W；骨骼肌；钙稳态；细胞凋亡

英文摘要： Chicken skeletal muscle injury induced by selenium deficiency could cause the content of Selenoprotein W decrease, calcium dyshomeostasis. The secondary structure of Selenoprotein W contains calmodulin domain. However, its mechanism of action of calcium homeostasis is not clear. This study established the model of selenoprotein W gene knock chicken and overexpression models of selenoprotein W gene knockout skeletal muscle cells. Base on the cases model of chicken muscle damage induced by selenium deficiency, using RNA interference, overexpression, flow cytometry, Western blot, real-time quantitative PCR and laser scanning confocal microscope to detect calcium homeostasis, content and mRNA level of calcium sensitivity receptor, calcium uptake and release of sarcoplasmic reticulum, calmodulin, calprotectin, muscle plasma nets related stress factor (GRP78/Bip、CHOP/GADDl53、XBP-1) and apoptosis, which could clarify the mechanism of Selenoprotein W on calcium homeostasis induced by selenium deficiency in chicken muscle damage. The implementation of this project will contribute to a better understanding of pathogenesis of selenium deficiency disease, but also to provide new ideas for comparative medicine in the field.

英文关键词： Chicken；selenoproteins W；skeletal muscle；calcium homeostasis；apoptosis