BDV核蛋白在少突胶质细胞内表达定位的基础研究  

Basic research of the expression and localization of BDV nucleoprotein in oligodendroglial cell

作　　者：王振海[1] 刘建[2] 谢鹏[3] 

机构地区：[1]宁夏医科大学附属医院神经内科,银川750004 [2]华北煤炭医学院附属医院神经内科,唐山063000 [3]重庆医科大学附属第一医院神经内科

出　　处：《中华微生物学和免疫学杂志》2010年第9期816-820,共5页Chinese Journal of Microbiology and Immunology

基　　金：2007年国家自然科学基金面上项目（30760067）;2007年宁夏自然科学基金项目（NZ0790）;2007年第四十二批中国博士后科学基金项目（20070420721）

摘　　要：目的 研究博尔纳病病毒核蛋白在转染的少突胶质细胞（oligodendroglial cell,OL）内的表达定位及其对细胞活性的影响.方法 应用RT-PCR检测转染细胞内BDVp40的基因片段,应用激光共聚焦显微镜和Western blot的方法观察核蛋白在细胞内的表达定位.应用MTT方法观察核蛋白对OL活性的影响.结果 在转染BDVp40基因片段的荧光表达质粒的OL内检测到BDVp40的基因片段;激光共聚焦显微镜提示核蛋白存在于细胞质和细胞膜中;Western blot的结果显示在细胞膜蛋白中存在核蛋白,而在细胞质中未检测到该蛋白;MTT法检测BDV核蛋白对OL活性具有明显的抑制作用.结论 通过研究转染的OL内稳定表达了BDV核蛋白,并将其定位在细胞的结构蛋白中,尤其在细胞膜中含量更为丰富,说明其可能在细胞的信号转导或介导病毒感染人胞过程中发挥关键作用.BDV核蛋白对OL的活性有抑制作用,这可能是BDV导致中枢神经系统损伤的机制之一.Objective To investigate the expression and location of Borna disease virus（BDV）nucleoprotein in transfected oligodendrocytes,and then to explore its expression mechanisms in oligodendroglial cell（OL）and their effects on cell proliferation.Methods We used PCR to detect BDVp40 gene fragments in transfected OL,laser confocal microscopy and Western blot method to detect the expression of nuclear protein and its intracellular location in the cell,MTT to detect the influence of cell proliferation by nuclear protein on the OL.Results We had detected the BDVp40 gene fragments in transfected oligodendrocytes.The nucleoprotein can be positioned in the cytoplasm and cell membrane by laser scanning confocal microscope;Western blot results showed that there is nucleoprotein in the constitutive protein,but was not detected in the cytoplasm.MTT tests showed that nucleoprotein expressed in OL can inhibit cell proliferation.Conclusion It is indicated that the transfected OL stablely express of BDV nucleoprotein,and itslocation is in the cell structure of proteins,particularly in the cytoplasm and more richer in cell membrane,that indicating they may play a key role in cell signal transduction or into the cell-mediated viral infection.BDV nucleoprotein inhibit proliferation of OL,which maybe an important mechanism of Borna disease virus persistant infection and produce symptoms.

关 键 词：博尔纳病病毒 核蛋白 少突胶质细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]