Quantitative Proteomics Analysis of Ischemia/Reperfusion Injury-Modulated Proteins in Cardiac Microvascular Endothelial Cells and the Protective Role of Tongxinluo
Quantitative Proteomics Analysis of Ischemia/Reperfusion Injury-Modulated Proteins in Cardiac Microvascular Endothelial Cells and the Protective Role of Tongxinluo
复制标题
心脏微血管内皮细胞缺血/再灌注损伤调节蛋白的定量蛋白质组学分析及通心络的保护作用
DOI：
10.1159/000470806
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发表时间：
2017-01-01
影响因子：
--
通讯作者：
Xu, Jun
中科院分区：
文献类型：
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作者：
Li, Qing;Cui, He-He;Xu, Jun
背景：内皮细胞（ECs）对再灌注损伤的保护作用很少受到关注。本研究采用串联质量标签（TMT）标记蛋白质组学方法，研究了缺血再灌注（I/R）损伤心肌微血管内皮细胞（CMECs）体外模型的调节蛋白及其中药通心络（TXL）的改变。方法：用不同浓度的TXL对人cmec进行缺氧2 h再氧2 h的处理，采用串联质谱法测定cmec的蛋白表达谱。我们评估了几种在I/R损伤中表达改变的蛋白，并总结了一些已报道的与I/R损伤相关的蛋白。结果：TXL呈剂量依赖性抑制CMEC细胞凋亡，最佳浓度为800µg/mL。I/R显著改变了cmec中的蛋白质，在正常组和缺氧及血清剥夺组之间检测到30种不同的蛋白质。在I/R损伤中，TXL上调6种蛋白，包括酰基辅酶A合成酶ACSM2B线粒体（ACSM2B）、细胞周期蛋白依赖性激酶抑制剂1B （CDKN1B）、血红素加氧酶1 （HMOX1）、转录因子SOX-17 （SOX17）、sequestoase -1异构体1 （SQSTM1）和TBC1结构域家族成员10B （TBC1D10B）。同时，TXL下调5种蛋白，包括血管生成素-2异构体c前体（ANGPT2）、细胞色素c氧化酶组装因子5 （COA5）、结缔组织生长因子前体（CTGF）、组织蛋白酶L1异构体2 （CTSL）和真核延伸因子2激酶（LOC101930123）。这类蛋白主要具有重要的功能，包括细胞增殖、应激反应和代谢过程的调节。结论：该研究通过蛋白质组学分析发现了I/R损伤后的差异蛋白。TXL调节cmes中蛋白的表达，在I/R反应中具有保护作用。
Background: The protection of endothelial cells (ECs) against reperfusion injury has received little attention. In this study, we used Tandem Mass Tag (TMT) labeling proteomics to investigate the modulated proteins in an in vitro model of cardiac microvascular endothelial cells (CMECs) subjected to ischemia/reperfusion (I/R) injury and their alteration by traditional Chinese medicine Tongxinluo (TXL). Methods: Human CMECs were subjected to 2 h of hypoxia followed by 2 h of reoxygenation with different concentrations of TXL Protein expression profiles of CMECs were determined using tandem mass spectrometry. We evaluated several proteins with altered expression in I/R injury and summarized some reported proteins related to I/R injury. Results: TXL dose-dependently decreased CMEC apoptosis, and the optimal concentration was 800 µg/mL. I/R significantly altered proteins in CMECs, and 30 different proteins were detected between a normal group and a hypoxia and serum deprivation group. In I/R injury, TXL treatment up-regulated 6 types of proteins including acyl-coenzyme A synthetase ACSM2B mitochondrial (ACSM2B), cyclin-dependent kinase inhibitor 1B (CDKN1B), heme oxygenase 1 (HMOX1), transcription factor SOX-17 (SOX17), sequestosome-1 isoform 1 (SQSTM1), and TBC1 domain family member 10B (TBC1D10B). Also, TXL down-regulated 5 proteins including angiopoietin-2 isoform c precursor (ANGPT2), cytochrome c oxidase assembly factor 5 (COA5), connective tissue growth factor precursor (CTGF), cathepsin L1 isoform 2 (CTSL), and eukaryotic elongation factor 2 kinase (LOC101930123). These types of proteins mainly had vital functions, including cell proliferation, stress response, and regulation of metabolic process. Conclusions: The study presented differential proteins upon I/R injury through a proteomic analysis. TXL modulated the expression of proteins in CMECs and has a protective role in response to I/R.