肝纤维化组织相关功能蛋白质组的差异分析

Differential analysis of associated proteome in hepatic fibrosis tissue

目的建立肝纤维化组织双向凝胶电泳图谱,初步分析蛋白质组的变化与差异表达。方法利用2-DE 分离肝纤维化组织和正常肝组织总蛋白质后,经图像分析识别差异表达的蛋白点,应用基质辅助激光解吸电离飞行时间质谱鉴定差异蛋白质,并用 Western 印迹方法对其中3个蛋白质的表达水平变化进行验证。结果比较分析肝纤维化组织和正常肝组织的2-DE 图谱,找到差异蛋白点59个,其中在肝纤维化组织表达上调30个,下调29个。并对其中15个表达差异3倍以上的蛋白点进行了肽质量指纹图分析,鉴定出10个与细胞信号转导、细胞增殖、氧化应激有关的蛋白质,14-3-3β、DJ-1及 PEBP 蛋白的 Western 印迹验证结果与2-DE 的检测结果相一致。结论肝纤维化组织和正常肝组织之间存在一些差异表达的蛋白质,为进一步阐明肝纤维化的发生机制奠定基础。

Objective To develop the 2-DE profiles of proteome from hepatic fibrosis tissue and preliminarily analyze the differential expression of proteome. Methods The differential expression of proteins were analyzed by imaging analysis and MALDI-TOF-MS after the total protein was extracted from the hepatic fibrosis tissue and the normal liver tissue by 2-DE. 3 proteins were verified by in Western-blotting. Results Fifty-nine differentially expressed protein were found in the proteome profile analysis of these two types of tissue, among which 30 protein spots were up-regulated and 29 protein spots were down-regulated in these hepatic fibrosis tissues. Fifteen protein spots with the differential expressions more than 3 times were analyzed by peptide mass fingerprint （PMF） and 10 differentially expressed proteins were found to be related with cell signal transduction, cell proliferation, and oxidative stress. Western-blotting results of 14-3-3β, DJ-1, and PEBP were consistent with those by the 2-DE examination. Conclusion Some differentially expressed proteins have been found in the hepatic fibrosis tissue and the normal liver tissue. These data provided a fundamental basis for further study of the mechanism of hepatic fibrogenesis.