Objective To evaluate the effect of this neuroendocrine hormone on protein expression by treating the human dermal fibroblasts with a-melanocyte stimulating hormone (α-MSH ).Methods Thehuman dermal fibroblasts was cultured, and the total protein of the fibroblasts were separated with immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE). After Coomassie bright blue staining, gel images were acquired by Image-scanner and then analyzed with the PDQuest software. 2-DE maps of fibroblasts were established. Partial differently expressed protein spots were incised from gels and digested by trypsin in-gel. Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and MSDB database searching by Mascot? software were used for protein identification. Results Well-resolved, reproducible 2-DE patterns of dermal fibroblasts treated with and without crMSH were obtained. 8 differently expressed protein spots were detected, among which 8 obtained peptide mass fingerprints (PMF) by MALDI-TOF-MS analysis. Among these proteins, of particular interest were five proteins annexin I, HSP27 and lamin A, etc. Conclusions Proteins expressed by human dermal fibroblasts treated with or without crMSH are different, and some of the differently expressed proteins involve apoptosis, intracellular signal transduction and framework construction and so on, which may be associated with anti-fibrosis effects of (a)-MSH on human dermal fibroblasts.%目的 观察α黑素细胞刺激素(α-melanocyte stimulating hormone,α-MSH)对人皮肤成纤维细胞作用的蛋白质表达谱的变化,从整体的蛋白质水平为α-MSH的抗纤维化机制研究提供线索.方法 体外培养人皮肤成纤维细胞,利用固相pH梯度(immobilized pH gradient,IPG)双向凝胶电泳(two-dimensional electrophoresis,2-DE)分离人皮肤成纤维细胞α-MSH处理组和对照组的总蛋白质,考马斯亮蓝显色,建立双向凝胶电泳图谱,经PDQuest软件图像分析,选取部分差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)分析获取肽质量指纹图谱,MascotTM软件搜索MSDB数据库鉴定蛋白质.结果 得到分辨率和重复性较好的α-MSH处理组和对照组人皮肤成纤维细胞2-DE图谱.筛选出8个差异表达蛋白质点进行质谱分析,经查询蛋白质序列数据库鉴定为膜联蛋白I、热休克蛋白27和核纤层蛋白A等.结论 人皮肤成纤维细胞在α-MSH作用后蛋白质的表达谱有差异,其差异表达的部分蛋白质功能涉及细胞凋亡、细胞内信号转导及细胞骨架构建等方面的因素,可能与α-MSH的抗纤维化作用有关.
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