皮肤老化过程中角质形成细胞的比较蛋白质组分析与鉴定

Analysis and identification of comparative proteome in keratinocytes druing skin aging process

目的筛选出表皮角质形成细胞中与皮肤老化相关的蛋白。方法从角质形成细胞(KC)中提取蛋白质,并进行2-DE分离,采用MALDI-TOF质谱分析及数据库查询;并在蛋白质水平进行Western-blotting验证。结果成功鉴定了21个差异表达蛋白,发现P21既随年龄增长高表达,也在同一年龄段曝光组高表达;而annexin A2的表达降低;在同一年龄段热休克蛋白质(HSP)27和HSP70、丝裂源激活的蛋白酶(MAPK)、Keratin,trpeⅡcyloskeletal 80(K2C80)在曝光部位高表达,而actin cyto-plasmic 1(ACTB)在非曝光部位高表达。并与蛋白质水平进行的验证结果一致。结论所鉴定的蛋白质与皮肤自然老化和(或)光老化关系密切,但其确切功能有待进一步研究。

Objective This study is to explore the differentially expressed protein or a specific protein expression that can be a marker of skin aging from an exposed and non-exposed skin of youth,middle-aged and elderly.Methods The proteins were extracted from the exposed and non-exposed elderly separated epidermis,and were submitted to two-dimensional gel electrophoresis.Then their electrophoretic patterns were analyzed by the PDQUEST software and identified by MALDI-TOF MS.The following steps were database query and probation using Western-Blotting.Results By comparison the two groups of two-dimensional electrophoresis patterns,in which 21 differential protein spots were successfully identified by MALDI-TOF analysis and database queries,and 4 proteins,HSP27、HSP70、MAPK、K2C80,were identified high expressed in exposed group while ACTB was high expressed in non-exposed group.Conclusion The changes in expression of 21 indetified proteins may have a close relationship with photoaging,that helps revealing molecular mechanism of epidermal cells in photoaging.