采用蛋白芯片结合SELDI—TOF—MS技术筛选紫外线照射后皮肤成纤维细胞损伤的特异性蛋白

Screening of specific proteins in ultraviolet-induced acute damage of human fibroblasts using proteinchip with surface enhanced laser desorption/ ionization time-of-flight mass spectrometry

目的探讨应用蛋白芯片结合表面增强激光解吸离子化飞行时间质谱（SELDI—TOF—MS）技术筛选紫外线照射后皮肤成纤维细胞早期损伤的生物标记的可行性。方法用弱阳离子交换蛋白芯片（CM10）结合SELDI-TOF-MS技术，检测未照射组、2．5J／cm^2。长波紫外线（UVA）照射组和100mJ／cm^2中波紫外线（UVB）照射组正常人真皮成纤维细胞差异表达的蛋白。结果紫外线照射组与未照射组蛋白质指纹图谱比较，发现10种蛋白表达下降，8种蛋白表达增强。将差异蛋白峰在SWISS—PROT和TREMBLE蛋白数据库中搜索，发现相对分子质量为11320．1的蛋白峰与人半胱氨酸天冬氨酸蛋白酶-7前驱体（caspase-7precursor）相匹配；相对分子质量为8574．3的蛋白峰与细胞色素C氧化酶多肽Vie前体（Cytochrome c oxidase polypeptide VIc precursor）相符。结论蛋白芯片结合SELDI—TOF-MS技术是一种快速、简便的高通量蛋白质组研究方法，为光损伤研究提供了新的研究平台。

Objective To investigate specific proteins in ultraviolet （UV）-induced acute damage of human fibroblasts using proteinchip with surface enhanced laser desorption/ionization time-of-flight mass spectrometry （SELDI-TOF-MS）. Methods CM10 Proteinchip and SELDI-TOF-MS were used for detecting protein samples of non-irradiated group, UVA （ 2. 5 J/cm^2 ）-irradiated group and UVB （ 100 mJ/cm^2 ） - irradiated group of normal human fibroblasts. Results Ten protein peaks were down-regulated and 8 protein peaks up-regulated in fibroblasts after UV irradiation. By searching in SWISS-PROT and TREMBLE databases, the peaks of 11 320. 1 and 8 574. 3 accorded with those of caspase-7 precursor and cytochrome C oxidase polypeptide VIc precursor proteins. Conclusion As a quick and convenient high-throughout proteomic analytic method, SELDI-TOF-MS plus proteinchip offers a unique platform for detecting dermal photodamage.