中文摘要：

目的初步探讨紫外线辐射所致皮肤细胞分子伴侣蛋白质组整体变化规律。方法分别提取30mJ／cm^2UVB照射前后角质形成细胞HaCaT株和10J／cm^2UVA照射前后成纤维细胞的总蛋白，采用固相pH梯度双向凝胶电泳技术进行分离，经Imaging Master 2D软件分析以发现差异表达蛋白，并对部分差异表达蛋白进行基质辅助激光解吸电离飞行时间（MALDI-TOF）质谱分析，测定其肽质量指纹图谱，在网上蛋白质数据库检索鉴定差异蛋白。采用Western免疫印迹法对部分质谱鉴定的差异蛋白做进一步验证。结果获得良好的双向电泳图谱，图像分析显示在紫外线辐射前后很多蛋白质发生差异表达，对差异表达的部分蛋白点进行肽质量指纹图谱分析，经Mascot软件检索人非冗余蛋白质数据库后，在UVB辐射后角质形成细胞中鉴定出3个表达显著增加的分子伴侣蛋白：热休克蛋白70KDa 9B、热休克蛋白60KDa和PHB；在UVA辐射后成纤维细胞中鉴定出4个表达显著增加的分子伴侣蛋白：热休克蛋白70KDa A5、热休克蛋白70KDa 9B、蛋白二硫键异构酶和PHB。Western免疫印迹法进一步验证上述结果。对这些蛋白的功能和与紫外线辐射的可能关系进行了初步探讨。结论紫外线辐射可以诱导皮肤细胞分子伴侣蛋白质组发生改变，而这些蛋白的变化可能与皮肤细胞应对紫外线辐射并清除其损伤相关。

英文摘要：

Objective To reveal the proteomic change of molecular chaperone in skin cell induced by UV radiation. Methods The keratinocyte （ HaCaT strain） was irradiated by 30mJ/cm^2 UVB ; and the fibroblast was irradiated by 10J/cm^2 UVA. The total proteins of these skin cells before and after UV radiation were separated by immobilized pH gradient based on two dimensional gel electrophoresis （2-DE）. The differential expression proteins were analyzed by using Imaging Master 2D analysis ware. The differential spots were identified by peptide mass fingerprinting based on Matrix - assisted laser desorption ionization time of flight mass spectrometry （MALDI-TOF-MS） and database searching were performed on the internet. Western blotting was used to further test the results of MALDI-TOF-MS. Results The 2D image was stable and showed lots of differential protein spots in skin cells after UV radiation. 3 molecular chaperone proteins which were up-expressed in keratinocyte after UVB radiation were identified ： HSP 70KDa 9B, HSP 60kDa, prohibitin ;4 molecular chaperone proteins which were up-expressed in fibroblast after UVA radiation were also identified ： HSP 70kDa A5, HSP 70KDa 9B, prohibitin, Disulfide isomerase. The results of western blotting were similar to those of MALDI-TOF-MS. Conclusion UV radiation could induce changes of the molecular chaperone expression pattern in keratinocyte and fibroblast. This proteinic changes could help skin cells to remove the impairment of UV radiation.