利用血清蛋白质组学技术筛选银屑病相关蛋白

Screening for psoriasis-associated proteins by serological proteome analysis

目的研究常见亚型银屑病患者血清中差异表达蛋白，筛选银屑病特异性蛋白质标记物。方法收集首次发病进展期寻常性银屑病6例、红皮病性银屑病5例及健康人6例的血清，分别将各组标本混合，并去除高丰度白蛋白和免疫球蛋白IgG后，应用双向电泳技术分离血清，比较差异蛋白点。利用基质辅助激光解析电离飞行时间质谱，对候选差异表达蛋白点进行肽质量指纹图谱分析，通过NCBI数据库搜索鉴定蛋白。结果获得了3组较好的血清双向电泳图谱。应用胶图分析软件分析，发现寻常性银屑病组、红皮病性银屑病组与健康对照组间差异蛋白点分别为33个和17个，寻常性银屑病组和红皮病性银屑病组之间差异蛋白点26个。共鉴定得到14种蛋白。在银屑病组中表达升高的有补体成分3、白介素16、维生素D结合蛋白、仅1抗胰蛋白酶等；红皮病组中补体成分3、补体因子H、仅1抗胰蛋白酶、血液结合素、触珠蛋白的表达量高于健康人对照，a1抗胰蛋白酶、补体因子H、补体成分4和触珠蛋白的表达量高于寻常性银屑病组。在红皮病性患者血清中表达下降的有血清淀粉样蛋白P。结论寻常性银屑病、红皮病性银屑病与健康人对照间血清蛋白表达谱存在差异。

Objective To screen for differentially expressed proteins in sera from patients with common types of psoriasis, and to identify plasma protein markers for psoriasis. Methods Serum samples were collected from 6 patients with progressive psoriasis vulgaris, 5 patients with erythroderma psoriaticum, and 6 healthy human controls, and then pooled into 3 pools： psoriasis vulgaris pool, erythroderma psoriaticum pool and control pool. After removal of high-abundance albumin and IgG, the pooled samples were analyzed by two-dimensional electrophoresis （2-DE）. An electrophoretic gel image analysis software was used to locate differentially expressed protein spots followed by peptide mass fingerprinting with matrix-assisted laser desorption ionization time of flight mass spectrometry （MALDI-TOF MS）. The National Center for Biotechnology Information （NCBI） protein databases were then searched for the identification of differentially expressed proteins. Results All the three pooled serum samples were well seperated by 2-DE. As the gel image analysis software showed, there were 33 protein spots differentially expressed between the patients with psoriasis vulgaris and the healthy controls, 17 between the patients with erythroderma psoriatieum and the healthy controls, and 26 between the patients with psoriasis vulgaris and those with erythroderma psoriaticum. Finally, 14 proteins were identified as differentially expressed proteins. The patients with psoriasis vulgaris showed higher expression of complement component 3, interleukin-16, vitamin D-binding protein and od-antitrypsin compared with the healthy controls; the patients with erythroderma psoriaticum showed increased expression of complement component 3, complement component H, a1-antitrypsin, hemopexin and haptoglobin, but decreased expression of serum amyloid protein compared with the healthy controls, as well as enhanced expression of a1-antitrypsin, complement component H, complement component 4 and haptoglobin compared with those with psoriasis vulgaris. Conclusion Differences exist in serum protein profiles between patients with psoriasis vulgaris and erythroderma psoriaticum, and healthy human controls.