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目的:筛选与人结肠上皮衰老相关的蛋白质以探讨人结肠上皮衰老及对癌症易感的分子机制。方法:应用双向凝胶电泳技术分离青年人及老年人的正常结肠上皮的总蛋白,经银染显色、图像分析、识别差异表达的蛋白质点,应用基质辅助激光解吸电离飞行时间质谱得到相应的肽质量指纹图谱,搜索数据库鉴定部分差异蛋白质点。结果:建立了青年人和老年人的正常结肠上皮的双向凝胶电泳图谱,质谱鉴定得到青年人和老年人之间的表达差异大于2倍的蛋白质点19个,包括17种非冗余蛋白质,它们的功能涉及物质代谢、能量产生、物质转运、抗氧化以及蛋白质的翻译、修饰和折叠等。结论:筛选并鉴定了17种结肠上皮衰老相关的蛋白质,提示线粒体功能受伤、抗氧化能力下降是结肠上皮衰老的重要原因,为揭示结肠上皮的衰老机制及衰老的结肠上皮对癌症易感的机制提供了线索。
OBJECTIVE: To screen the proteins related to human colon epithelial senescence to explore the molecular mechanism of human colon epithelial senescence and its susceptibility to cancer. Methods: The total protein of normal colorectal epithelium of young and old people was separated by two-dimensional gel electrophoresis. The differentially expressed protein spots were identified by silver staining, image analysis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry Peptide mass fingerprinting, search database identified partial differential protein spots. Results: The two-dimensional gel electrophoresis pattern of normal colonic epithelium was established in young people and old people. Mass spectrometry identified 19 protein spots with more than 2-fold difference in expression between young and old, including 17 kinds of non-redundant proteins, Their function involves material metabolism, energy production, substance transport, antioxidant and protein translation, modification and folding. CONCLUSION: Seventeen kinds of proteins related to aging of colon epithelium were screened and identified, which implies that mitochondrial function is impaired and the anti-oxidative capacity is reduced. It is an important reason for the aging of colonic epithelium. To reveal the mechanism of aging of colon epithelium and the susceptibility mechanism of aging colon epithelium to cancer Provided clues