HCV患者外周血单个核细胞蛋白质表达质谱分析

目的:应用比较蛋白质组学的方法分析丙型肝炎患者外周血单个核细胞蛋白质表达模式的变化及寻找丙型肝炎相关生物标记分子,进一步识别鉴定其差异表达蛋白质,分析其对丙型肝炎慢性化机制的意义.方法:应用固相化pH梯度双向凝胶电泳(2-DE)分离健康者(10)及HCV患者(28)PBMC的总蛋白质,凝胶银染显色后,PDQuest图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)获得差异蛋白点的肽质指纹图谱,通过SWISS-PROT数据库鉴定蛋白质.结果:得到两张2-DE图谱,HCV患者及健康者PBMC凝胶的蛋白质点数分别为625及614;初步筛选出HCV患者与健康者存在明显差异的12个蛋白点,经质谱分析,初步鉴定了10种蛋白质.这些差异蛋白质包括病毒蛋白、蛋白质合成与分解、三大代谢相关酶类、细胞结构相关蛋白质以及信号转导相关蛋白质.结论:应用2-DE及MALDI-TOF-MS方法建立了HCV患者PBMC双向凝胶电泳图谱,分离并初步鉴定了10种与HCV感染相关的差异表达的蛋白质,为研究HCV慢性化相关机制提供新的线索.

Mass spectrometric analysis of peripheral blood mononuclear cells in HCV infected patients

AIM: To investigate the diversity of proteins' pattern of the peripheral blood mononuclear cells (PBMCs) in patients with hepatitis C virus (HCV) infection, identify the differentially expressed proteins and analyze their roles in the mechanism of chronic hepatitis C. METHODS: The total proteins from PBMCs in HCV infected patients (28) and healthy persons (10) were separated by the immobilized pH gradientjbased 2-DE. The differentially expressed proteins were screened by PDQuest analysis software, and then identified by peptide mass fingerprint (PMF) based on matrixjassisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and database searching. RESULTS: 2-DE results showed that the mean number of protein spots in HCV infected patients and healthy persons was 625 and 614.Twelve differential proteins were tested by MALDI-TOF-MS, and 10 of them were identified. Some of the proteins participated in protein synthesis and degradation, signal transduction, metabolism, or cytoskeleton construction while some of them were proteins of virus. CONCLUSION: 2-DE pattern of PBMCs from HCV infected patients or healthy persons has been established and ten differentially expressed proteins has been characterized. Our study is helpful for clinical detection of HCV infection and further research into the mechanisms of chronic hepatitis C.