几丁糖对人成纤维细胞蛋白质表达谱的影响

Effects of chitosan on human fibroblasts ： a proteomic analysis

目的观察几丁糖对人成纤维细胞蛋白质表达谱的影响。方法取第5代人成纤维细胞，实验组用含1g／L几丁糖的培养液作用24h，对照组仅用培养液培养24h。分别提取实验组和对照组的成纤维细胞总蛋白，用双向凝胶电泳进行分离，银染后进行图像分析，识别有表达差异的蛋白质点；用基质辅助激光解析电离飞行时间串联质谱技术鉴定部分差异蛋白质点。结果实验组和对照组有43个差异蛋白质点，其中17个点在实验组中表达上调，26个点在实验组中表达下调。经质谱分析，鉴定出6个已知蛋白，3个未知蛋白。已知蛋白分别与细胞凋亡、信号转导、细胞骨架及代谢等有关。结论在几丁糖作用下，成纤维细胞的部分蛋白质表达发生变化。

Objective To observe the effect of chitosan on protein maps of human fibroblasts. Methods The human fibroblasts at the 15th generation were divided into experimental and control groups. The cells in experimental group were cultured in DMEM with 1 g/L chitosan for 24 h, and those in control group in DMEM only. The total proteins of both groups were extracted, and separated by two-dimensional gel electrophoresis （2-DE）. After colloidal silver staining, the images were analyzed by using Imagemaster 5.0 software to discriminate protein spots with differential expression. The distinct spots were picked and digested, then identified by peptide mass fingerprints （PMF） and database searching. Results Atlas of 2-DE were successfully established. There were 43 differentially expressed spots, 17 of which were higher and 26 lower in experimental group. By PMF analysis ,6 known proteins were identified. These proteins were involved in apoptosis, signal transduction, cytoskeleton, metabolism, etc. Conclusion After treatment with chitosan, the protein profile of fibroblasts has changed.