非标记定量技术研究特应性皮炎患者血清的差异蛋白

【摘要】 目的 应用非标记定量技术研究特应性皮炎患者血清中的差异表达蛋白，筛选特异性蛋白质标记物。 方法 收集特应性皮炎患者和健康对照者的血清各6份，处理和提取血清蛋白质，经一维SDS-PAGE和蛋白胶内酶解获得肽段，应用高效液相色谱串联质谱鉴定，使用Mascot软件查询并用Scaffold软件作相对定量分析。 结果 特应性皮炎患者组和健康对照组的血清共检测出76种差异表达超过1.5倍的蛋白质，患者组血清表达上调的有50种蛋白质，表达下调的有26种蛋白质，其中有26种蛋白质仅在患者组表达，14种蛋白质仅在对照组表达。 结论 特应性皮炎患者与健康人血清间存在蛋白质表达差异。

Application of label-free quantitative proteomics in the screening for differentially expressed serum proteins in patients with atopic dermatitis

Liu Sujun, Xu Jian, Zhao Bin, Zhu Huijun, Sun Yan, Xu Jin, Shen Hong. Department of Dermatology, Third People′s Hospital of Hangzhou, Hangzhou 310009, ChinaCorresponding author：Shen Hong, Email: shenhangzhou@sina.com 【Abstract】 Objective To screen differentially expressed serum proteins by label-free quantitative proteomics in patients with atopic dermatitis, in order to discover serum protein markers for atopic dermatitis. Methods Serum samples were collected from 6 patients with atopic dermatitis and 6 healthy human controls. Then, the samples from the patients were pooled together, so with those from the controls. Proteins were extracted from the two joint serum samples followed by separation with one-dimensional sodium dodecyl sulfate polyacrylamide gel electropheresis （SDS-PAGE）. Peptides were obtained by enzymolysis in protein gels, which were subsequently identified by high performance liquid chromatography tandem mass spectrometry. The Mascot software was used for database searching, and the Scaffold software for relatively quantitative analysis of proteins. Results Totally, 76 proteins with a difference of greater than 1.5 folds were identified between the serum samples of patients with atopic dermatitis and controls. There were 50 up-regulated and 26 down-regulated serum proteins in the patients compared with the healthy controls. Among these differentially expressed proteins, 26 were only detected in the sera of the patients, and 14 proteins in those of the controls. Conlusion Differences exist in the expression of serum proteins between atopic dermatitis patients and healthy people．