基于iTRAQ技术的卒中后抑郁症患者血浆蛋白质组学研究

发布时间：2018-05-29 20:39

本文选题：卒中后抑郁症 + 中风　；参考：《重庆医科大学》2014年硕士论文

【摘要】：研究背景脑卒中后抑郁是脑卒中患者中最常见的精神疾病并发症。卒中后抑郁可以影响到患者脑卒中的治疗效果，加重身体残疾，干扰患者的肢体功能恢复，增加患者的痛苦，甚至使患者产生自杀观念。但是，目前卒中后抑郁疾病发生发展潜在的病理生理机制尚不清楚，在临床上也没有可用于疾病诊断或病情监测的检测方法。目的本研究旨在运用蛋白质组学技术探讨卒中后抑郁疾病患者血浆蛋白质组的变化，拟寻找卒中后抑郁患者血浆中异常表达的蛋白质，以期作为客观诊断卒中后抑郁症的生物标志物，从而为疾病的诊断、预防、发病机制及治疗提供依据。方法本研究是对卒中后抑郁组（15例）、卒中组（15例）和正常对照组（15例）受试者的血浆蛋白质进行的一个比较蛋白质组学研究。首先对三组受试者的血浆进行14种高丰度蛋白的去除，，然后采用iTRAQ同位素标记与多维液相色谱分离串联质谱技术对三组受试者的样本进行相对定量分析和质谱鉴定，最后通过免疫印迹法对蛋白质组学结果进行进一步验证，并对显著改变的差异蛋白搜索数据库进行生物信息分析。结果蛋白质组学研究结果显示蛋白表达量显著改变的蛋白质的生物学功能主要与脂质代谢和免疫调节有关，我们选择了与这些生物功能密切相关的6个蛋白（ApoA-IV，ApoC-II，CRP，gelsolin,haptoglobin，LRG）进行了Western blotting验证。与卒中组比较，蛋白ApoA-IV的表达量在卒中后抑郁中显著上调，ApoC-II、 LRG和CRP三个蛋白的表达量在卒中后抑郁组和正常对照组中显著下调。而gelsolin和haptoglobin这两个蛋白的表达量在三组中都有显著地改变（gelsolin：正常对照组卒中后抑郁组卒中组；haptoglobin,卒中组卒中后抑郁组正常对照组）。结论由于gelsolin和haptoglobin这两个蛋白显示出独特的蛋白分层表达谱，所以gelsolin蛋白表达量的提高联合haptoglobin蛋白表达量的降低可能可以作为临床上卒中患者产生抑郁症状风险增加的诊断标志物
[Abstract]:Research background Post-stroke depression is the most common complication of mental illness in stroke patients. Post-stroke depression can affect the therapeutic effect of stroke, aggravate the body disability, interfere with the recovery of limb function, increase the pain of patients, and even lead to suicidal concept. However, the underlying pathophysiological mechanism of post-stroke depression is not clear, and there is no clinical detection method for disease diagnosis or disease monitoring. Purpose The aim of this study was to investigate the changes of plasma proteome in patients with post-stroke depression by using proteomics techniques, and to seek abnormal expression of proteins in plasma of patients with post-stroke depression. It is expected to be used as a biomarker for the objective diagnosis of post-stroke depression and to provide evidence for the diagnosis, prevention, pathogenesis and treatment of the disease. Method This study was a comparative proteomic study of plasma proteins in 15 patients with post-stroke depression, 15 patients with stroke and 15 normal controls. At first, 14 kinds of high abundance proteins were removed from the plasma of the three groups, and then the samples of the three groups were analyzed quantitatively by iTRAQ isotope labeling and tandem mass spectrometry with multi-dimensional liquid chromatography. Finally, the proteomics results were further verified by Western blotting, and the significant change of differential protein search database was analyzed by bioinformatics. Result The results of proteomics showed that the biological functions of proteins with significant changes in protein expression were mainly related to lipid metabolism and immunomodulation. We selected six proteins, ApoA-IVApoC-II, CRPgelsolinhaptoglobinine (LRGG), which were closely related to these biological functions, to be verified by Western blotting. Compared with stroke group, the expression of protein ApoA-IV significantly up-regulated the expression of ApoC-II, LRG and CRP in post-stroke depression group and normal control group. However, the expression of gelsolin and haptoglobin were significantly changed in the three groups: normal control group, post-stroke depression group, stroke group, normal control group. Conclusion Because the two proteins gelsolin and haptoglobin show unique protein layering expression profile, the increase of gelsolin protein expression combined with the decrease of haptoglobin protein expression may be used as a diagnostic marker for the increased risk of depression in patients with stroke.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743.3;R749.4

【参考文献】