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[稳定型心绞痛血浆生物标志物的筛选与验证:差异蛋白质组学分析]

[Screening and verification of plasma biomarkers for stable angina pectoris: a differential proteomic analysis].

作者信息

Shen An-Na, Hu Shui-Wang, Huang Jing, Zheng De-Zhong, Hu Zhao-Ting

机构信息

Department of Cardiovascular Medicine, 3Department of Clinical Laboratory, Third Affiliated Hospital, Southern Medical University, Guangzhou 510630, China. E-mail:

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2017 Oct 20;37(10):1370-1374. doi: 10.3969/j.issn.1673-4254.2017.10.14.

Abstract

OBJECTIVE

To compare and analyze the differentially expressed plasma proteome between patients with stable angina pectoris (SAP) and healthy donors to identify the biomarkers for early diagnosis of SAP.

METHODS

Plasma samples from 60 patients with SAP and 60 healthy controls were collected. Twenty samples (100 mL each) randomly selected from each group were pooled and after removing high-abundance proteins from the pooled plasma, two-dimensional gel electrophoresis (2DE) was performed to isolate the total proteins. The protein spots with more than 2 fold changes were selected after 2D analysis using software, and the differentially expressed proteins were identified by MALDI TOF/TOF mass spectrometer. ELISA was performed to detect hemoglobin subunit delta (HBD) levels in 40 randomly selected samples from each group for verification of the results of 2DE.

RESULTS

A total of 7 differentially expressed proteins were found in plasma samples from patients with SAP, including 3 up regulated proteins (serum albumin, hemoglobin subunit alpha and hemoglobin subunit delta,) and 4 down?regulated ones (apolipoprotein L1, apolipoprotein C3, apolipoprotein E and complement C4B). ELISA results showed that HBD level was increased in SAP plasma, which was consistent with the results of 2DE.

CONCLUSION

Patients with SAP have different plasma protein profiles from those of healthy controls, and HBD may serve as a potential specific biomarker for early diagnosis of SAP.

摘要

目的

比较和分析稳定型心绞痛(SAP)患者与健康供体之间差异表达的血浆蛋白质组,以鉴定SAP早期诊断的生物标志物。

方法

收集60例SAP患者和60例健康对照者的血浆样本。从每组中随机选取20个样本(每个100 mL)混合,在去除混合血浆中的高丰度蛋白质后,进行二维凝胶电泳(2DE)分离总蛋白。使用软件进行二维分析后,选择变化倍数超过2倍的蛋白点,并用基质辅助激光解吸电离飞行时间/飞行时间质谱仪鉴定差异表达的蛋白质。对每组随机选取的40个样本进行酶联免疫吸附测定(ELISA)以检测血红蛋白亚基δ(HBD)水平,以验证2DE的结果。

结果

在SAP患者的血浆样本中总共发现7种差异表达的蛋白质,包括3种上调蛋白(血清白蛋白、血红蛋白亚基α和血红蛋白亚基δ)和4种下调蛋白(载脂蛋白L1、载脂蛋白C3、载脂蛋白E和补体C4B)。ELISA结果显示,SAP血浆中HBD水平升高,这与2DE的结果一致。

结论

SAP患者与健康对照者的血浆蛋白质谱不同,HBD可能作为SAP早期诊断的潜在特异性生物标志物。

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本文引用的文献

1
CLINICAL PRACTICE. Chronic Stable Angina.临床实践。慢性稳定性心绞痛。
N Engl J Med. 2016 Mar 24;374(12):1167-76. doi: 10.1056/NEJMcp1502240.
2
The importance of hemoglobin A2 determination.血红蛋白A2测定的重要性。
Rev Bras Hematol Hemoter. 2015 Sep-Oct;37(5):287-9. doi: 10.1016/j.bjhh.2015.06.002. Epub 2015 Jul 21.
4
Hemoglobin α in the blood vessel wall.血管壁中的血红蛋白α
Free Radic Biol Med. 2014 Aug;73:136-42. doi: 10.1016/j.freeradbiomed.2014.04.019. Epub 2014 May 14.

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