microRNA-155在鸡缺硒性脾淋巴细胞凋亡中的作用机制研究

项目名称： microRNA-155在鸡缺硒性脾淋巴细胞凋亡中的作用机制研究

项目编号： No.31472161

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 李术

作者单位： 东北农业大学

项目金额： 83万元

中文摘要： 脾淋巴细胞过度凋亡是鸡缺硒性免疫抑制机制之一，microRNA-155参与调控免疫器官的分化、发育和细胞凋亡，但其在鸡缺硒性免疫器官脾脏的细胞凋亡中的作用尚不清楚。本课题拟在建立鸡缺硒性脾脏淋巴细胞凋亡模型的基础上，应用实时荧光定量PCR、免疫印迹、透射电镜和组学技术等，检测脾脏转录组、蛋白组和microRNA-155的表达，结合生物信息学技术，筛选microRNA-155的靶基因及其在鸡缺硒性脾脏细胞凋亡发生中的可能信号转导通路，在此基础上，体外培养鸡脾脏淋巴细胞，建立microRNA-155抑制与过表达模型、其靶基因敲低和过表达模型，应用流式细胞术、激光共聚焦、实时定量荧光PCR和免疫印迹等方法，检测细胞凋亡、钙稳态、靶基因表达等，阐明microRNA-155在鸡缺硒性脾脏细胞凋亡发生中的信号转导通路，为防治缺硒性免疫功能障碍提供理论依据，也为比较医学提供借鉴。

中文关键词： microRNA-155；硒；鸡；脾淋巴细胞；细胞凋亡

英文摘要： Excessive apoptosis in spleen lymphocyte is the main mechanism of immunosuppressive induced by selenium deficiency in chicken. MicroRNA-155 involved in regulating the immune organ differentiation, development and apoptosis. However, the role of microRNA-155 in apoptosis in chicken spleen lymphocyte still unclear.The main reason of immunosuppression induced by selenium deficiency in chicken is spleen lymphocyte apoptosis. MicroRNA-155 is the major one of microRNA in immune system, which it not only plays an important role in the development and differentiation of immune organ, but also influence cell apoptosis through the special signal transduction pathways. Base on the cases model of chicken splenic lymphocyte apoptosis induced by selenium deficiency in vitro, using real-time quantitative PCR, western blot, transmission electron microscope and omics technology methods to detect the transcriptome, proteomics and the level of microRNA-155. Combined with bioinformatics technology, screening the target genes of microRNA-155 and the possible signal pathway in chicken splenic lymphocytes induced by selenium deficiency. On this basis, modeling microRNA-155 knockdown and overexpression splenic lymphocytes in vitro, using flow cytometry, laser confocal microscopy, real-time quantitative PCR and western blot methods to detect the cell apoptosis, calcium homeostasis and the expression of target genes. The implementation of this project will contribute to a better understanding of the mechanism of microRNA-155 on the signal transduction in cell apoptosis in chicken splenic lymphocytes. The result of this project will provide not only a basis of prevention and treatment of immune function barrier induced by selenium deficiency, but also a theoretical reference for comparative medicine.

英文关键词： microRNA-155;selenium;chicken;splenic lymphocytes;apoptosis