Two-Dimensional Fluorescence In-Gel Electrophoresis of Coronary Restenosis Tissues in Minipigs Increased Adipocyte Fatty Acid Binding Protein Induces Reactive Oxygen Species-Mediated Growth and Migration in Smooth Muscle Cells
Two-Dimensional Fluorescence In-Gel Electrophoresis of Coronary Restenosis Tissues in Minipigs Increased Adipocyte Fatty Acid Binding Protein Induces Reactive Oxygen Species-Mediated Growth and Migration in Smooth Muscle Cells
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DOI：
10.1161/atvbaha.112.301016
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发表时间：
2013-03-01
影响因子：
8.7
通讯作者：
Shen, Wei Feng
中科院分区：
文献类型：
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作者：
Lu, Lin;Wang, Ya Nan;Shen, Wei Feng
我们的目的是通过定量二维荧光凝胶电泳（2D-DIGE）揭示有和没有链脲佐菌素诱导的糖尿病的小型猪冠状动脉支架内再狭窄（ISR）组织的蛋白质变化，并研究鉴定的关键蛋白质，特别是脂肪细胞脂肪酸结合蛋白（AFABP），方法和结果：西罗莫司洗脱支架植入15只糖尿病和26只非糖尿病小型猪的冠状动脉，6个月后重复血管造影。通过2D-DIGE和MALDI-TOF/TOF质谱分析糖尿病和非糖尿病小型猪中重要ISR和非ISR节段的内膜组织。糖尿病和非糖尿病小型猪ISR组织中AFABP水平均显著高于非ISR组织，且糖尿病ISR组织中AFABP水平高于非糖尿病ISR组织。在人动脉平滑肌细胞中，AFABP的过度表达显著改变了表型，促进了生长和迁移，在高葡萄糖中的作用比在低葡萄糖培养基中更突出，而AFABP敲低抑制了这些作用。AFABP过表达通过多种途径上调NADPH氧化酶亚基Nox 1、Nox 4和P22的表达，增加活性氧的产生，同时升高下游基因细胞周期蛋白D1、基质金属蛋白酶2和单核细胞趋化蛋白1。然而，AFABP诱导的作用被抑制diphenyleneiodonium，通路抑制剂，和小干扰RNA。此外，从AFABP表达的人动脉平滑肌细胞和重组AFABP的上清液也促进细胞的生长和migration.Conclusion-This研究表明，AFABP显着增加在冠状动脉ISR段的糖尿病和非糖尿病小型猪。人动脉平滑肌细胞AFABP表达增加和分泌AFABP通过活性氧介导的活化促进生长和迁移（Arterioscler Thromb Vasc Biol.2013;33：572-580.）
Objective-We aimed to uncover the protein changes of coronary artery in-stent restenosis (ISR) tissue in minipigs with and without streptozotocin-induced diabetes mellitus by quantitative 2-dimensional fluorescence in-gel electrophoresis (2D-DIGE), and to investigate the influences of crucial proteins identified, particularly adipocyte fatty acid binding protein (AFABP), in human arterial smooth muscle cells.Methods and Results-Sirolimus-eluting stents were implanted in the coronary arteries of 15 diabetic and 26 nondiabetic minipigs, and angiography was repeated after 6 months. The intima tissue of significant ISR and non-ISR segments in both diabetic and nondiabetic minipigs was analyzed by 2D-DIGE and MALDI-TOF/TOF mass spectrometry. AFABP level was significantly increased in ISR tissue than in non-ISR tissue in both diabetic and nondiabetic minipigs, with level being higher in diabetic ISR than in nondiabetic ISR tissue. In human arterial smooth muscle cells, overexpression of AFABP significantly altered phenotype and promoted growth and migration, with effects more prominent in high-glucose than in low-glucose medium, whereas AFABP knockdown inhibited these effects. AFABP overexpression increased reactive oxygen species production by upregulating the expression of NADPH oxidase subunits Nox1, Nox4, and P22 through multiple pathways, with elevation of downstream gene cyclin D1, matrix metalloproteinase-2, and monocyte chemoattractant protein-1. However, AFABP-induced effects were inhibited by diphenyleneiodonium, pathway inhibitors, and small interfering RNA. In addition, the supernatant from AFABP-expressing human arterial smooth muscle cells and recombinant AFABP also promoted cellular growth and migration.Conclusion-This study has demonstrated that AFABP is significantly increased in coronary artery ISR segments of both diabetic and nondiabetic minipigs. Increased AFABP expression and secretory AFABP of human arterial smooth muscle cells promote growth and migration via reactive oxygen species-mediated activation. (Arterioscler Thromb Vasc Biol. 2013;33:572-580.)