UVA辐射前后成纤维细胞蛋白质组双向电泳图谱的差异分析

Differential Analysis of Two-dimensional Gel Electrophoresis Profiles of Fibroblast Proteome Before and After UVA Radiation

目的使用双向电泳(2-DE)比较UVA辐射前后成纤维细胞蛋白质的差异,从分子水平探索紫外线辐射所致皮肤细胞蛋白整体变化的规律。方法采用固相pH梯度双向凝胶电泳分离UVA辐射前后人皮肤成纤维细胞的总蛋白质,凝胶经银染显色后,用Im agingM aster 2D图像分析软件进行比较分析,识别差异表达的蛋白质。结果①UVA辐射前后人皮肤成纤维细胞凝胶的平均蛋白质点数分别为676±21和693±28,平均匹配的点数分别为583.43±29.13和591.46±27.23,匹配率达86.22%和85.31%。②UVA辐射前后人皮肤成纤维细胞的平均匹配蛋白点数为5 6 3.4 7±19.33。差异表达蛋白点数为17个,其中1个点仅在UVA辐射前成纤维细胞中表达,另4个点仅在UVA辐射后成纤维细胞中表达;7个点在UVA辐射后成纤维细胞中高表达,5个点在UVA辐射后成纤维细胞中低表达。结论本研究建立了分辨率高且重复性较好的UVA辐射前后成纤维细胞的双向凝胶电泳图谱,发现两者间存在一些差异表达的蛋白质,以上差异点的发现为研究紫外线对皮肤作用的机理提供了有益的线索。

Objective To compare the proteomic change of fibroblast induced by UVA radiation. Methods The total proteins of fibroblast before and after UVA radiation were separated by immobilized pH gradient based two dimensional gel electrophoresis （2-DE） . After silver staining,the differential expression proteins were analyzed by using Imaging Master 2D analysis ware. Results ①Average protein spots were 676 ± 21,693 ± 28 in fibroblast before and after UVA radiation respectively, the matched spots were5 83.43 ± 29. 1 3 , 59 1 . 46 ± 27. 23 in both the two types of tissues respectively, and the average matching rate was 86.22% ,85.31% respectively. ②A total of 563.47 ± 19.33 protein spots was matched between the electrophoretogram of fibroblast before and after UVA radiation. The 17 differential protein spots were identified between the two types of the tissues. 1 protein spot was expressed only in the cells before UVA radiation and 4 protein spots were expressed only in the fibroblast after UVA radiation. The expression levels of 7 protein spots in the fibroblast after UVA radiation were higher than those before UVA radiation, and the expression levels of 5 protein spots in the fibroblast after UVA radiation were lower than those before UVA radiation. Conclusion In this study, the well-resolved, reproducible 2-DE patterns of fibroblast before and after UVA radiation were established and some differential proteins between the two types of tissues were found. These data will be helpful to the further study of UV effect on skin.