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目的:利用蛋白质组学方法,识别并鉴定幽门螺杆菌(Helicobacterpylori,H.pylori)处理HepG2细胞后差异表达的蛋白质,为进一步探讨幽门螺杆菌对肝细胞的病理作用机制奠定基础。方法:运用固相pH梯度双向凝胶电泳技术分离H.pylori处理前后HepG2细胞的总蛋白质,用图象分析软件比较分析,以识别差异表达的蛋白质;应用质谱仪(massspectrometry)得到相应的肽质指纹图谱(peptidemassfingerprint,PMF),然后搜索数据库鉴定差异蛋白质。结果:鉴定出12种差异表达蛋白质,这些蛋白质包括整合素β1、蛋白激酶Cα、PINCH蛋白、Ras相关蛋白Rab37、LIM同源盒蛋白Lhx1、HLAⅠ类抗原、血管抑制素和金属蛋白酶抑制因子2等。这些差异蛋白质涉及基因表达调控、细胞免疫、细胞生长和信号传导等众多关键事件。结论:H.pylori处理前后HepG2的蛋白质组具有差异,这些差异表达的蛋白质可能参与了H.pylori对肝细胞的病理作用过程,这种蛋白质组的差异分析有助于进一步研究H.pylori与人类肝脏疾病的关系。
OBJECTIVE: To identify and identify the differentially expressed proteins of HepG2 cells by Helicobacter pylori (H.pylori) proteomics method, and lay a foundation for further exploration of the mechanism of H. pylori on the pathology of hepatocytes. Methods: The total proteins of HepG2 cells before and after H.pylori treatment were isolated by solid-phase pH gradient two-dimensional gel electrophoresis and compared with image analysis software to identify differentially expressed proteins. Mass spectrometry (mass spectrometry) Fingerprinting (peptidemassfingerprint, PMF), and then search the database to identify differential proteins. RESULTS: Twelve differentially expressed proteins were identified. These proteins include integrin β1, protein kinase Cα, PINCH, Ras-related protein Rab37, LIM homeobox protein Lhx1, HLA class I antigens, angiostatin and metalloproteinase 2 . These differential proteins are involved in many key events such as gene expression regulation, cellular immunity, cell growth and signaling. Conclusion: There are differences in HepG2 proteome before and after H.pylori treatment. These differentially expressed proteins may be involved in the pathological process of H.pylori on hepatocytes. The differential analysis of this proteome will be helpful to further study the relationship between H.pylori and human Liver disease.