博尔纳病病毒感染对少突胶质瘤细胞中IL-1β表达量的影响  

Effects of Borna disease virus infection on the expression of IL-1β protein of oligodendrocyte

作　　者：张中海[1] 车洋[2] 

机构地区：[1]徐州医学院生理学教研室,江苏徐州221004 [2]哈尔滨医科大学病原微生物学教研室,黑龙江哈尔滨150086

出　　处：《徐州医学院学报》2009年第4期230-232,共3页Acta Academiae Medicinae Xuzhou

基　　金：黑龙江省教育厅科学技术研究项目(11511159);黑龙江省教育厅骨干教师创新能力项目;哈尔滨市科技攻关计划项目(2005AA9CS116 -16)

摘　　要：目的探讨博尔纳病病毒(Borna disease virus,BDV)感染对少突胶质瘤细胞(oligodendrocyte,OL)中白细胞介素1β(interleukin 1β,IL-1β)蛋白表达的影响。方法首先将OL和BDV感染的OL煮沸裂解,分别提取蛋白混合液,测定蛋白的浓度后,将二者的蛋白混合液浓度均调至10 g/L,分别取蛋白混合液及其经10 000 r/min离心5 min后的上清,沉淀后分别进行斑点免疫印迹试验(dot-blot hybridization),检测OL和BDV感染的OL中IL-1β的存在;同时,对提取的OL和BDV感染的OL中的蛋白混合液进行2倍系列稀释后进行斑点免疫印迹试验,检测细胞中的IL-1β的表达量的变化。结果斑点免疫印迹试验表明,OL和BDV感染的OL中提取的蛋白混合液及收集的沉淀均出现杂交阳性信号,而蛋白混合液离心后的上清未出现杂交阳性信号。等量的OL和BDV感染的OL的蛋白混合液经2倍系列稀释后进行斑点免疫印迹试验,OL的蛋白混合液在≤32倍的稀释度范围内有杂交阳性信号,BDV感染的OL的蛋白混合液在≤8倍的稀释度范围内有杂交阳性信号。结论①IL-1β在OL和BDV感染的OL中均存在,且存在于本实验获得的蛋白混合液和沉淀中。②BDV感染使OL中的IL-1β的表达量降低,提示IL-1β的表达量下降,有可能使其参与的脑内的免疫生理及病理反应发生变化,这可能是BDV感染致病的机制之一。Objective To investigate the effects of Borna disease virus （BDV） infection on the expression of inter- leukin 1β（ IL - 1β） protein of oligodendroeytes （OL）. Methods Both OL and the BDV - infected OL were subjected to boiling lysis, and their protein mixture was respectively extracted. After concentration determination, the protein mixtures were adjusted to 10 g/L and centrifuged at 10 000 r/min for 5 rain to extract protein mixtures and supematant, which were precipitated to have dot - blot hybridization, respectively for determination of IL - 1β in OL and BDV - infected OL. Meanwhile, the protein mixtures extracted from OL and BDV - infected OL were tested by dot - blot hybridization after 1 ： 2 serial dilution to determine the variations of the expression levels of IL - 1β in the cells. Results Dot - blot hybridization showed that positive hybrid signals were present in both the protein mixtures and precipitates extracted from OL and BDV -infected OL, while no positive hybrid signals were detected in the supernatant obtained from centrifuged protein mixtures. Equal amount of protein mixtures of the OL and the BDV - infected OL were tested by dot - blot hybridization after 1 ： 2 serial dilution. Positive hybrid signals were detectable in protein mixtures of OL within the range ≤ 1 ： 32 serial dilution, while in the protein mixtures of BDV - infected OL positive hybrid signals were detectable within the range ≤1 ： 8. Conclusion （1）IL - 1 β are present in both OL and BDV - infected OL, and in this experiment, present in the protein mixtures and precipitates. （2）BDV infection leads to the decreased expression of IL - 1β, suggesting that the decrease in the level of IL - 1β expression might be involved in the changes of immunological, physiological and pathological reactions within the brain and might be one of the pathogenic mechanisms of BDV infection.

关 键 词：博尔纳病病毒 少突胶质瘤细胞 白细胞介素-1Β 斑点免疫印迹试验 

分 类 号：R446.62[医药卫生—诊断学]