Chen Cao, Xiao Di, Zhou Wei, Shi Qi, Zhang Hui-Fang, Zhang Jin, Tian Chan, Zhang Jian-Zhong, Dong Xiao-Ping
State Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center of Infectious Disease, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, People's Republic of China.
Mol Neurobiol. 2014 Feb;49(1):290-302. doi: 10.1007/s12035-013-8519-2. Epub 2013 Aug 4.
The shotgun proteomic based on the approach of tandem mass tag (TMT) labeling has received increasing attention for neuroproteomics analysis and becomes an effective tool for the identification and quantification of a large number of proteins for the purpose of revealing key proteins involved in the neuronal dysfunction and an inflammatory response associated with neurodegenerative disorders. To assess the potential expression difference of proteins in cerebrospinal fluids (CSF) between Creutzfeldt-Jakob disease (CJD) and non-CJD patients, the pooled CSF samples from 39 Chinese probable sporadic CJD (sCJD) patients and from 52 non-CJD cases were comparably analyzed with the methodology of TMT labeling and RP-RP-UPLC-MS/MS. Totally, 437 possible proteins were identified in the tested CSF specimen, among them, 49 proteins with 95 % confidence interval. Differential assays showed among those 49 CSF proteins, 12 were upregulated and 13 were downregulated significantly in the sCJD compared to non-CJD. The most affected pathway of the differential expression proteins in CSF of sCJD was complement and coagulation cascade. Western blots for six selected changed proteins in the pooled CSF samples revealed the similar altering profiles in the groups of sCJD and non-CJD as proteomics. Furthermore, CSF samples from 24 CJD patients and 24 non-CJD patients were randomly selected and subjected individually into the Western blots of an increased protein (phosphoglycerate mutase 1) and a decreased one (alpha-1-antichymotrysin), which also confirmed the altering tendency of these identified proteins. Those data indicate that proteomic assay of CSF is a powerful technique not only for selection of the potential biomarkers for the development of diagnostic tool of CJD but also for supplement of useful scientific clues for understanding the CSF homeostasis during the pathogenesis of prion diseases.
基于串联质量标签（TMT）标记方法的鸟枪法蛋白质组学在神经蛋白质组学分析中受到越来越多的关注，并成为一种有效的工具，用于鉴定和定量大量蛋白质，以揭示与神经退行性疾病相关的神经元功能障碍和炎症反应中涉及的关键蛋白质。为了评估克雅氏病（CJD）患者与非CJD患者脑脊液（CSF）中蛋白质的潜在表达差异，采用TMT标记和反相-反相-超高效液相色谱-串联质谱法，对39例中国散发性CJD（sCJD）患者和52例非CJD患者的脑脊液混合样本进行了比较分析。在测试的脑脊液样本中总共鉴定出437种可能的蛋白质，其中49种蛋白质的置信区间为95%。差异分析显示，在这49种脑脊液蛋白质中，与非CJD相比，sCJD中有12种上调，13种下调。sCJD患者脑脊液中差异表达蛋白质受影响最大的途径是补体和凝血级联反应。对混合脑脊液样本中6种选定的变化蛋白质进行的蛋白质印迹分析显示，sCJD组和非CJD组的变化模式与蛋白质组学相似。此外，随机选择24例CJD患者和24例非CJD患者的脑脊液样本，分别对一种上调蛋白质（磷酸甘油酸变位酶1）和一种下调蛋白质（α-1-抗糜蛋白酶）进行蛋白质印迹分析，这也证实了这些鉴定蛋白质的变化趋势。这些数据表明，脑脊液蛋白质组学分析不仅是一种强大的技术，可用于选择CJD诊断工具开发的潜在生物标志物，还可为理解朊病毒疾病发病机制期间的脑脊液稳态提供有用的科学线索。
