帕金森病患者血浆蛋白质组学的初步分析及鉴定

Analysis and identification of plasma proteomics in patients with Parkinson's disease

目的分析帕金森病(PD)患者血浆蛋白质组学的变化并筛选PD临床诊断和干预相关的生物学标志物。方法选择2016年7月至2017年3月于新乡医学院第一附属医院神经内科就诊的PD患者33例,按照疾病分期将患者分为早期PD组(n=19)和晚期PD组(n=14);另选取同期36例健康体检者作为对照组。分别从3组受试者中随机选取6例受试者的血样进行蛋白质组学分析。利用高效液相色谱仪(HPLC)联合Q-Exactive质谱仪筛选出早期PD组、晚期PD组、对照组样本中的蛋白质表达谱,并统计差异表达的蛋白质分子。采用酶联免疫吸附试验(ELISA)检测早期PD组、晚期PD组和对照组受试者血浆中载脂蛋白E(APOE)、过氧化物酶蛋白-2(PRDX-2)、补体成分7(C7)蛋白表达水平。结果早期PD组与对照组之间筛选出97个差异性表达的蛋白质分子,晚期PD组与对照组之间筛选出68个差异性表达的蛋白质分子。差异表达的蛋白质生物信息学分析结果显示,这些蛋白质功能主要与氧化应激反应、炎症反应、补体激活、神经突触再生等生物学过程密切相关。3组受试者血浆中C7蛋白水平比较差异均无统计学意义(P>0.05)。早期PD组、晚期PD组患者血浆中APOE、PRDX-2蛋白水平均低于对照组(P<0.05)。结论 PD患者血浆中的蛋白质表达发生改变,APOE和PRDX-2可能是早期诊断PD的生物学标志物。

Objective To analyse the changes of plasma proteomics in patients with Parkinson’ s disease（ PD） and screen the biological markers which related to the clinical diagnosis and intervention of PD. Methods Thirty-three patients with PD in the Department of Neurology,the First Affiliated Hospital of Xinxiang Medical University from July 2016 to March 2017 were selected and divided into early-stage PD group（ n = 19） and advanced-stage PD group（ n = 14） according to the stage of disease. Thirty-six healthy persons were selected as control group. Six subjects were randomly selected from each group to perform the plasma proteomics analysis. The expression profile of plasma proteome in early-stage PD group,advanced-stage PD group and control group was screened by high-performance liquid chromatography（ HPLC） and Q-Exactive mass spectrometers,and the differentially expressed protein molecules were counted. The apolipoprotein E（ APOE）,peroxiredoxin-2（ PRDX-2）and complement component 7（ C7） protein expression levels were detected by enzyme linked immunosorbent assay. Results A total of 97 differentially expressed proteins were screened between the control group and early-stage PD group,and 68 differentially expressed proteins were screened between the control group and advanced-stage PD group. Bioinformatics analysis results showed that the function of these differentially expressed proteins were mainly related to the oxidative stress,inflammatory response,complement activation,and neuron projection regeneration. There was no statistic difference in the C7 protein expression level in plasma among the control group,early-stage PD group and advanced-stage PD group（ P 〉 0. 05）. The levels of APOE,PRDX-2 protein in early-stage PD group and advanced-stage PD group were significantly lower than those in the control group（ P 〈 0. 05）. Conclusion The protein expression in the plasma of PD patients is changed,and the APOE and PRDX-2 may be the biomarkers for the early diagnosis of PD.