三氯乙烯药疹样皮炎差异血清蛋白的筛选及鉴定

Screening and identification of differential serum proteins related to dermatitis medicamentosa-like of trichloroethylene

目的 筛选并鉴定三氯乙烯（trichloroethylene,TCE）暴露者、TCE药疹样皮炎急性期及恢复期差异血清蛋白.方法 收集TCE暴露者血清（Ⅰ组）、TCE药疹样皮炎急性期患者血清（Ⅱ组）和TCE药疹样皮炎恢复期患者血清（Ⅲ组）各5份,各组血清样本等量混合,去除白蛋白、IgG等高丰度蛋白后经除盐处理并定量.采用双向凝胶电泳技术对上述3组混合血清分别进行蛋白分离,银染后扫描成像,用ImageMaster Platinum 2D 5.0软件进行分析,筛选差异蛋白,并应用基质辅助激光解析电离飞行时间质谱（MALDI-TOF-MS）进行鉴定.结果 经去除高丰度蛋白和除盐处理后获得了分辨率高、重复性好的双向电泳图谱,处理后血清蛋白检出量增至（300±12）个.经MALDI-TOF-MS鉴定得到5个差异蛋白,分别为：补体4b、载脂蛋白A-Ⅰ、载脂蛋白C-Ⅲ、载脂蛋白C-Ⅱ和转甲状腺素蛋白.补体4bⅢ组相对Ⅰ组表达量为1.352 88,表达上调.Ⅱ组相对Ⅰ组血清中载脂蛋白C-Ⅱ相对表达量为1.512 14,表达上调;载脂蛋白A-Ⅰ、载脂蛋白C-Ⅲ和转甲状腺素蛋白相对表达量分别为-1.601 17、-1.034 49、-1.313 35,表达有所下调.结论 筛选的差异蛋白功能与机体免疫及肝功能异常有关,为阐明TCE致TCE药疹样皮炎作用机制及筛选相关生物标志物提供了重要信息.

Objective To screen and identify differential serum proteins which might be involved in dermatitis medicamentosa-like of trichloroethylene （DMLT）.Methods Three groups of sera were collected from population exposed to trichloroethylene（TCE）（group Ⅰ）,patients suffering from DMLT（group Ⅱ）,and the healed cases （group Ⅲ）.After removing albumin and IgG in the three pools of sera,a comparative proteomic analysis was carried out. The images were analyzed using ImageMaster Platinum 2D 5.0 to screen the differentially expressed proteins.The protein spots were then subjected to matrix-assisted laser desorption/ionization time-of flight mass spectrometry （MALDI-TOF-MS） and tandem mass spectrometry sequencing of tryptic peptides for further identification.Results The depletion of albaumin and IgG greatly increased the number of protein spots to 300±12.Five differential spots wore identifled.which were complement component C4b,apolipoprotein A-Ⅰ,apolipoprotein C-Ⅲ apolipoprotein C-Ⅱ and tranthyretin Compared with group Ⅰ,the expression levels of complement component C4b in group Ⅲ and apolipuprotein C-Ⅱ in group Ⅱ were up-regulated（1.352 88-fold,1.512 14-fold,respectively）;compared with group Ⅰ,the expression levels of apolipoprotein A-Ⅰ,apolipoprotein C-Ⅲ and transthyretin in group Ⅱ were down-regulated （1.601 17-fold,1.034 49-fold,1.313 35-fold,respectively）.Conclusion The findings of this study show that most of the identified differential proteins are closely related to immunity and liver dysfunction.which provides some evidence on elucidating the mechanisms and screening of biomarkers of TCE intoxication.