烟雾病相关血清蛋白标志物筛选及鉴定

Screening and identification of moyamoya disease-related serum protein biomarkers

目的筛选烟雾病血清差异蛋白,并研究其在烟雾病中的作用,为可能的烟雾病分子靶向治疗提供依据。方法于2015年12月—2017年3月分别收集烟雾病病人血清11份与正常人血清20份,去除Ig G、白蛋白等高丰度蛋白,胰蛋白酶酶切后i TRAQ标记,液相分离后质谱检测,筛选血清差异蛋白。筛选出TGF-β1后进行酶联免疫吸附试验(ELISA)验证。结果总共鉴定了202个烟雾病相关的血清差异蛋白,其中13个蛋白上调,7个蛋白下调,ELISA结果验证了维生素D结合蛋白(D6RF35)在烟雾病病人血清中表达水平异常升高(1.22±0.48,P<0.05),辅酶Q10B(Co Q10B)血清中含量明显降低(0.81±0.25,P<0.05);工作特征曲线(ROC)表明COQ10B曲线下面积(AUC)(AUC=71.4%)优于D6RF35(AUC=61.7%)。结论 COQ10B是烟雾病人潜在的血清标志蛋白,在其发病中可能起到关键作用,效果优于D6RF35。

Objective To screen moyamoya disease-related serum proteins and to explore associations of the protein with the disease for providing evidences to the development of molecular targeted therapy. Methods Serum samples were collected from 11 moyamoya disease patients and 20 healthy controls. The samples were labeled with isobaric tags for relative or absolute quantitation（i TRAQ） after the elimination of abundant proteins such as immunoglobulin G（Ig G） and albumin and trypsin digestion; then differential proteins in the samples were detected with liquid chromatography-tandem mass spectrometry（LC-MS/MS）. The identified transforming growth factor β1（TGF-β1） was verified with enzyme-linked immunosorbent assay（ELISA）. Results Among the 202 moyamoya disease-related serum proteins identified, 13 were upregulated and 7 were down-regulated. Significantly increased vitamin D-binding protein（D6 RF35）（1.22 ± 0.48 ng/ml） and decreased coenzyme Q10 B（Co Q10 B）（0.81 ± 0.25 ng/ml） were detected in the serum samples of moyamoya disease patients（both P〈0.05）. The receiver operating characteristic（ROC） curve analysis demonstrated a larger area under the ROC curve（AUC）（71.4%） of the Co Q10 B than that of D6 RF35（61.7%）. Conclusion Co Q10 may be a potential serum protein marker better than D6 RF35 for moyamoya disease and may play an important role in the incidence of moyamoya disease.