【摘要】 目的:建立用于脑脊液(CSF)蛋白分离的双向电泳(2-DE)技术体系,寻找多发性硬化患者和对照组脑脊液蛋白质差异,从分子水平探讨多发性硬化患者脑脊液蛋白整体变化规律。方法:分别取对照组及多发性硬化患者的脑脊液,用三氯乙酸(TCA)沉淀蛋白质后,加入适量裂解液使蛋白质充分裂解,离心后取上清液上样。进行第1向等电聚焦(IEF)电泳和第2向十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE),染色脱色后,分析所得蛋白质斑点。结果:对照组和多发性硬化患者脑脊液2-DE图谱上均可得到近200个蛋白质斑点,其中2个蛋白质在多发性硬化患者脑脊液中含量增加,4个蛋白质在多发性硬化患者脑脊液中含量减少,12个蛋白质斑点为多发性硬化患者脑脊液特有。结论:对照组和多发性硬化患者脑脊液的双向电泳图谱存在明显差别,这些发现可为研究多发性硬化疾病机制提供线索。更多还原

【Abstract】 Objective: To establish 2-dimensional electrophoresis system for cerebrospinal fluid(CSF), and to find differences of CSF proteins of healthy people and patients with multiple sclerosis(MS). Methods: CSF were extracted from patients with MS and controls, and the proteins were precipitated with trichloroacetic acid(TCA). Considerable lysis solution was added in for complete lysis of proteins, and then the superior solution was got after centrifugation. Isoelectric focusing(IEF) electrophoresis was employed as the first dimension and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) was employed as the second dimension. After being stained and bleached, the protein spots were analyzed. Results: Totally, about 200 spots were obtained in 2-DE maps of patients with MS and controls, respectively, of which 2 spots increased and 4 spots decreased in quantity in MS. Furthermore, 12 protein spots were specific to CSF of MS. Conclusion: There are distinctive differences in 2-DE maps of patients with MS and controls, which offers clues for study on machanisms of MS.更多还原