不稳定型心绞痛患者血浆蛋白标记物的蛋白质组学筛选

发布时间：2018-04-24 11:55

本文选题：心绞痛 + 不稳定型　；参考：《解放军医学杂志》2017年06期

【摘要】：目的分析冠心病不稳定型心绞痛(UAP)与稳定型心绞痛(SAP)患者血浆中的差异蛋白质,筛选可能与UAP早期诊断密切相关的血浆蛋白标记物。方法分别收集2014年6月-2015年4月南方医科大学第三附属医院UAP及SAP血浆标本各60例,另收集体检科收集的血浆标本作为正常对照组(n=60)。随机取对照组(n=10)、UAP组(n=10)与SAP组(n=10)空腹血浆标本各100μl,分别等量混合成3组样本,去除血浆高丰度蛋白后,利用双向差异凝胶电泳(DIGE)技术进行蛋白分离,经差异软件分析后,采集UAP和SAP之间变化2倍以上的差异蛋白质点,利用基质辅助激光解吸电离-飞行时间/飞行时间质谱(MALDI-TOF/TOF MS)对差异蛋白点进行鉴定。每组随机选取40份血浆样本,选取UAP特异性差异蛋白进行ELISA验证。结果 UAP与SAP组患者血浆相比较,共筛选出10个表达量差异2倍以上的差异蛋白点,包括9个上调蛋白点,1个下调蛋白点。经质谱鉴定后,表达上调的蛋白包括纤维蛋白原γ链(FGG)、补体C4-B(C4B)、免疫球蛋白κ链C结构域(IGKC)和血红蛋白α亚基(HBA1);表达下调的蛋白是结合珠蛋白(HP)。与对照组比较后,在这些差异蛋白中共找到2个UAP特异性相关蛋白,即IGKC和HP。选取IGKC进行ELISA验证,结果表明,与对照组和SAP组相比较,UAP组样本中IGKC特异性表达上调(P0.05),与DIGE验证结果一致。结论筛选到UAP特异性相关蛋白IGKC和HP,IGKC有可能成为UAP早期筛查及诊断的特异性生物标记物。
[Abstract]:Objective to analyze the differential proteins in the plasma of patients with unstable angina pectoris (UAP) and stable angina pectoris (SAP) and to screen the plasma protein markers which may be closely related to the early diagnosis of UAP. Methods the plasma samples of UAP and SAP were collected from June 2014 to April 2015 in the third affiliated Hospital of Southern Medical University, respectively. The plasma samples collected from the Department of physical examination were taken as the normal control group. The fasting plasma samples of the control group and the SAP group were randomly mixed into 3 groups in the same volume. After removing the plasma high abundance protein, the protein was separated by two-dimensional differential gel electrophoresis (DIGE), and the protein was analyzed by differential software. The differential protein spots were collected and identified by matrix assisted laser desorption ionization / time of flight mass spectrometry (MALDI-TOF / TOF MS). 40 plasma samples were randomly selected in each group and UAP specific differential protein was selected for ELISA verification. Results 10 differentially expressed protein spots, including 9 up-regulated protein spots and 1 down-regulated protein point, were screened out in UAP group compared with those in SAP group. The up-regulated proteins were identified by mass spectrometry, including fibrinogen 纬 chain FGGGG, complement C4-BnC4BN, immunoglobulin 魏 chain C domain IGKC) and hemoglobin 伪 subunit HBA1C, and the down-regulated protein was the binding globin (HPN). Compared with the control group, two UAP specific related proteins, IGKC and IGKC, were found in these differentially expressed proteins. IGKC was selected for ELISA validation. The results showed that the specific expression of IGKC was up-regulated in UAP group compared with control group and SAP group, which was consistent with the result of DIGE verification. Conclusion the screening of UAP specific related proteins IGKC and HPG-IGKC may be specific biomarkers for early screening and diagnosis of UAP.
【作者单位】：南方医科大学病理生理学教研室、广东省蛋白质组学重点实验室;南方医科大学第三附属医院心血管内科;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

【参考文献】