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目的分析病毒性脑炎(VE)患儿和对照组儿童脑脊液(CSF)蛋白质组的表达差异,筛选VE特异性蛋白,探讨其与VE的关系,为VE的早期诊断提供线索。方法以三氯醋酸/丙酮沉淀法提取VE患儿和对照组儿童CSF总蛋白,固相pH梯度二维聚丙烯酰胺凝胶电泳(2-DE)技术分离蛋白质,应用双向电泳凝胶分析软件(PDQuest 7.3.1)对2-DE凝胶进行量化比较分析,识别差异表达明显的蛋白质点,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)得到肽质量指纹图谱(PMF),在Mascot数据库中进行蛋白质匹配,鉴定部分差异蛋白质。结果VE患儿和对照组CSF蛋白2-DE凝胶图谱上分别平均显示442和401个点,有23个点蛋白质表达存在2倍以上的差异,选取5个表达明显上调的蛋白质点进行质谱鉴定,成功鉴定出钙结合蛋白S100B。结论钙结合蛋白S100B在VE患儿和无中枢神经系统病变的对照组儿童CSF的蛋白表达上存在明显差异,可能是VE密切相关的疾病特异性蛋白,并有可能成为VE诊断的分子标志物。
Objective To analyze the differential expression of cerebrospinal fluid (CSF) proteome in children with viral encephalitis (VE) and control children, screen VE-specific proteins and investigate their relationship with VE, and provide clues for the early diagnosis of VE. Methods The total protein of CSF was extracted from children with VE and control group by trichloroacetic acid / acetone precipitation method. The proteins were separated by two-dimensional polyacrylamide gel electrophoresis (2-DE) PDQuest 7.3.1) were used to quantify and compare the 2-DE gel to identify differentially expressed protein spots. Peptide mass fingerprinting (PMF) was performed using matrix-assisted laser desorption / ionization time of flight mass spectrometry (MALDI-TOF-MS) Mascot database for protein matching, identify some of the differential proteins. Results On the 2-DE gel map of CSF, there were 442 and 401 spots on average in the VE children and control group respectively. There were more than 2 times of protein expression in 23 spots. Five spots with significantly up-regulated protein spots were identified by mass spectrometry , Successfully identified calcium binding protein S100B. Conclusions There is a significant difference in the protein expression of calcium-binding protein S100B between children with VE and control group without central nervous system disease, which may be closely related disease-specific proteins and may become a molecular marker for the diagnosis of VE.