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.docVIP

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 【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10000 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 IL-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。BDV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。 【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 　　Abstract: Objective To investigate the effects of Borna disease virus (BDV) infection on the expression of interleukin 1β (IL-1β) protEin of oligodendrocytes (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 10000 r/min for 5 min to extract protein mixtures and supernatant, 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