增生性瘢痕表皮细胞差异蛋白的初步分析

Differentially expressed proteins of hypertrophic scar epidermal cells

目的检测并鉴定增生性瘢痕角质形成细胞(HK)较正常者差异表达的蛋白质,揭示 HK在增生性瘢痕(HS)形成过程中的作用。方法利用消化法分离5例HS(均位于四肢、处在增生期)组织HK,提取其中总蛋白质,利用双向电泳技术展示蛋白质分子的差异表达,并对差异蛋白进行质谱分析。结果初步建立了>600个点的在体HK双向电泳图谱,筛选差异蛋白质点24 个,其中HK较正常高表达点8个,低表达点9个,极低表达4个,新发现的蛋白质点3个;对2个点质谱鉴定结果为Maspin前体与Zinc finger protein 226。结论消化分离法获得在体HK并建立其2-D图谱的方法可行,为HS相关蛋白质组研究奠定了一定基础。初步筛选的24个差异蛋白质点及质谱分析所得结果提示HK蛋白表达存在异常,可能与HS的形成有关。

Objective To explore the function of the keratinocy-tes during the course of the formation of hypertrophic scar （HS）. Methods The keratinocy-tes of 5 proliferating HS tissues from extremities were isolated with Dispase Ⅱ and trypsin. Total proteins of the cells were extracted and separated. The differential expression of proteins was displayed by two-directional electrophoresis and the differentiaUy expressed proteins were analyzed by using mass spectrogram. Results The final optimized 2-directional map of more than 600 protein spots and 24 differentially expressed proteins were detected by the software, including 8 up-regulated ones, 13 down-regulated ones in HK and 3 newly identified ones. Two proteins named Maspin precursor and Zinc finger protein 226 were identified by mass spectrogram. Condusion It is feasible to establish the noncultured HK 2-directial map which made the further study on hypertrophic scar proteomics possible, The 24 differentially expressed proteins might play a role in the formation of hypertrophic scar.