稳定型心绞痛患者血浆差异蛋白的筛选及验证

发布时间：2018-01-29 03:01

本文关键词： 稳定型心绞痛血浆生物标记物蛋白质组学双向凝胶电泳　出处：《南方医科大学学报》2017年10期 　论文类型：期刊论文

【摘要】：目的分析稳定型心绞痛(SAP)患者与健康对照组血浆中差异表达的蛋白质组,筛选可能与SAP诊断密切相关的血浆生物标记物。方法采集南方医科大学第三附属医院SAP血浆标本60例,另收集体检科收集的血浆标本作为正常对照组(n=60)。下面不变,做凝胶电泳是将各组的血混合的做实验的。取SAP患者组与健康对照组空腹血浆标本各20例(各100 mL),分别等量混合成2组样本,去除血浆高丰度蛋白后,利用双向凝胶电泳技术进行蛋白分离,经差异软件分析后,挖取变化2倍以上的差异蛋白质点,利用基质辅助激光解析电离-飞行时间/飞行时间质谱对差异蛋白点进行鉴定。每组随机各取40例样本,选取差异蛋白血红蛋白δ亚基进行酶联免疫吸附实验验证。结果 SAP患者与健康人血浆相比较,筛选出表达量差异2倍以上的差异蛋白共7种,表达上调的蛋白包括血清白蛋白、血红蛋白α亚基和血红蛋白δ亚基;表达下调的蛋白包括载脂蛋白L1、载脂蛋白C3、载脂蛋白E和补体C4-B。ELISA验证结果表明血红蛋白δ亚基在SAP组样本中表达上调,与双向凝胶电泳结果一致。结论血浆血红蛋白δ亚基有可能成为稳定型心绞痛早期筛查及诊断的特异性生物标记物。
[Abstract]:Objective to analyze the differential expression of proteome in plasma between patients with stable angina pectoris (SAP) and healthy controls. Methods 60 plasma samples of SAP from the third affiliated Hospital of Southern Medical University were collected. In addition, the plasma samples collected by the Department of physical examination were taken as the normal control group. The fasting plasma samples were collected from 20 patients with SAP and 20 healthy controls (100ml / L each), and were mixed into two groups of samples in the same quantity. After removing the plasma high abundance protein, the protein was separated by two-dimensional gel electrophoresis. After the difference software analysis, the difference protein spots with more than 2 times change were extracted. The differential protein sites were identified by matrix assisted laser desorption ionization / time of flight mass spectrometry. 40 samples were randomly collected from each group. The differential protein hemoglobin 未 subunit was selected to be verified by enzyme-linked immunosorbent assay (Elisa). Results compared with the healthy plasma, 7 differentially expressed proteins were obtained in patients with SAP. The up-regulated proteins included serum albumin, hemoglobin 伪 subunit and hemoglobin 未 subunit. The down-regulated proteins including apolipoprotein L1, apolipoprotein C3, apolipoprotein E and complement C4-B.ELISA showed that hemoglobin 未 subunit was up-regulated in SAP group. Conclusion Plasma hemoglobin 未 subunit may be a specific biomarker for early screening and diagnosis of stable angina pectoris.
【作者单位】：南方医科大学第三附属医院心血管内科;南方医科大学病理生理学教研室;南方医科大学第三附属医院检验科;
【基金】：广东省科技计划项目(2013B021800309)
【分类号】：R541.4
【正文快照】： 冠心病已经成为威胁全人类的第1大死亡病因[1]。研究表明,约半数以上的冠心病患者出现心绞痛(AP)的临床症状[2]。研究显示SAP病人平均年死亡率仅为2%~3%[3],在临床上并未引起足够的重视。然而,有些SAP患者也同样会进展为急性冠脉综合征,危害性更加严重。目前,对于SAP的临床诊

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