调节猪免疫功能的关键硒蛋白的探寻及其表达的调控机制

研究证实，营养水平或稍高于营养水平的硒具有显著调节猪免疫特别是细胞免疫的作用。硒蛋白是硒在体内发挥生物学功能的主要形式，已确定的硒蛋白有20余种。但究竟哪些硒蛋白在猪免疫调节中起关键作用？硒蛋白的表达是如何调控的？尚未见研究。本项目主要是通过猪原代淋巴细胞培养和仔猪试验，应用CFSE示踪和流式细胞术等方法检测T淋巴细胞增殖（用抗猪CD3单抗等刺激）、ELISA检测细胞因子、荧光定量PCR和Western Blot检测硒蛋白表达并结合RNA干扰技术，研究硒、猪免疫功能与硒蛋白基因表达或沉默的相互关系，探寻调节猪免疫功能的关键硒蛋白；应用RNA干扰和基因芯片技术研究上调或下调硒蛋白基因的表达会导致哪些相关基因发生变化，筛选与硒蛋白基因相互作用的基因及可能的信号通路。目的是阐明硒调节猪免疫的分子机理，为新型硒源饲料添加剂的研制提供标示硒蛋白，同时为硒蛋白的开发利用（如新型免疫佐剂）提供科学依据。

It has been confirmed that selenium at the nutritional levels or slightly higher than the nutritional levels plays an important role in the regulation of immunity in pigs, especially in the cellular immunity. The physiological functions of selenium are largely due to the presence of selenoproteins. Now more than 20 selenoproteins have been identified, but it's still not clear which selenoproteins play a key role in the regulation of immunity in pigs and how the selenoprotein expression are modulated by selenium status. The present project will investigate the effects of selenium on the pig immune function and its relationships to the selenoprotein expressions and seek the key selenoproteins on the regulation of immunity induced by selenium by using CFSE labeling and flow cytometry (evaluating T lymphocytes proliferation stimulated with anti-pig CD3 mAb), ELISA (cytokines assay), Real-time PCR, Western Blot （seleniprotein expression assay）and RNAi technologies in vitro and in vivo. The changes of some related genes after up or down-regulation of the selenoprotein expressions and the possible involved signaling pathways also will be studies by using RNAi and gene chip technologies. The aim of this project is to elucidate the molecular mechanism of immune regulation induced by selenium in pigs, to give an evidence for the application of some specific selenoproteins as a biomarker in development of a new selenium-source additive and to provide a scientific basis for the application of selenoproteins on animal husbandry (e.g. a new immuno-adjuvant).