基于蛋白组学技术筛选急性主动脉夹层诊断标记物

Screening diagnostic biomarkers of acute aortic dissection by proteomics

目的 应用非标记定量蛋白组学比较急性主动脉夹层（AAD）与急性心肌梗死（AMI）和健康人血清中的差异蛋白,筛选并验证潜在的夹层早期诊断标志物.方法 前期预实验收集急性主动脉夹层患者与健康人血清各3份,进行非标记蛋白组学检测,筛选出差异蛋白.之后收集急性主动脉夹层、急性冠脉综合征患者和健康人血清各9份,对目标蛋白进行Elisa验证,同时收集夹层与正常动脉组织进行免疫组化验证.结果 谱图计数（spectral counts）共鉴定出648种蛋白,其中夹层组较健康人组升高4倍的蛋白有12种.对其中升高明显的纽蛋白（vinculin）进行Elisa验证,夹层组[n=9,（1366.95±394.85）pg/ml]与心梗组[n=9,（2656.91 ±531.98） pg/ml]、健康人组[n=9,（3949.37±314.61）pg/ml] （P＜0.05）相比并没有升高;免疫组化发现主动脉发生撕裂部位的纽蛋白（vinculin）分布较未发生撕裂部位密集.结论 非标记定量蛋白质组学技术是寻找AAD循环标志物的一种有效手段.纽蛋白可能参与AAD的发病过程,是一种具有研究价值的AAD潜在循环标志物.

Objective To screen and identify potential serum biomarkers for acute aortic dissection using label-free proteomics. Methods Serum samples were collected from 3 AAD patients and 3 healthy people. Label free quantitation proteomic approach was used to identify the proteins in serum. The proteins with significant differ- ence are validated as the candidate proteins （fold changes〉4 or 〈0.25, P〈0.05）. Then collected serum samples from 9 AAD patients and 9 AMI patients and 9 healthy controls were detected for target protein by Elisa, and AAD and normal aortic tissues were collected to verify the location of target protein. Results Among 648 types of proteins which were identified by proteomic, 12 proteins were significantly different（fold changes〉4 or 〈0.25, P〈 0.05）. The protein, vinculin, was significantly higher than the other proteins. The serum levels of vinculin by ELISA showed that the AAD group was lower than the AMI group and the healthy group [ （1366.95±394.85）pg/ml vs （2656.91 ±531.98 ）pg/ml vs （3949.37 ±314.61 ）pg/ml, P〈0.05 ] though immunohistochemistry staining demon- strated that vinculin distributed more intensively at the laceration site. Conclusion Our study indicates that label- free quantitation proteomics may be an available method for finding circulation biomarkers for AAD diagnosis. Vin- culin, which is probably involved in the pathogenesis of AAD, is a potential circulation biomarker for AAD.