Advanced glycation end products upregulate lysyl oxidase and endothelin-1 in human aortic endothelial cells via parallel activation of ERK1/2-NF-κB and JNK-AP-1 signaling pathways
Advanced glycation end products upregulate lysyl oxidase and endothelin-1 in human aortic endothelial cells via parallel activation of ERK1/2-NF-κB and JNK-AP-1 signaling pathways
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DOI：
10.1007/s00018-015-2091-z
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发表时间：
2016-04-01
影响因子：
8
通讯作者：
Papavassiliou, Athanasios G.
中科院分区：
文献类型：
--
作者：
Adamopoulos, Christos;Piperi, Christina;Papavassiliou, Athanasios G.
内皮功能障碍涉及关键细胞外基质 (ECM) 酶赖氨酰氧化酶 (LOX) 和血管收缩蛋白内皮素-1 (ET-1) 的失调，其基因表达可由转录激活因子核因子 kappa B (NF-kappa B) 和激活蛋白-1 (AP-1) 调节。晚期糖基化终末产物 (AGE) 是内皮功能障碍的加重因素，其与受体 RAGE 结合后会诱导丝裂原激活蛋白激酶 (MAPK) 上调，从而导致 NF-κ B 和 AP-1 增强。我们假设 AGE 可以通过 AGE/RAGE/MAPK 信号轴诱导 LOX 和 ET-1 表达的 NF-kappa I' 和 AP-1 依赖性调节。在用 AGE-牛血清白蛋白 (AGE-BSA) 处理后的人主动脉内皮细胞 (HAEC) 中采用蛋白质印迹、实时 qRT-PCR、FACS 分析和电泳迁移率变化分析来研究这一假设的信号传导途径。此外，对暴露于高或低 AGE 含量饮食的大鼠实验模型的主动脉内皮进行 AGE、RAGE、LOX 和 ET-1 表达的免疫组织化学分析。在不同时间点暴露于 AGE-BSA 的 HAEC 表现出 LOX 和 ET-1 mRNA 水平以剂量和时间依赖性方式上调。 HAEC 暴露于 AGE-BSA 还显示出磷酸 (p)-ERK1/2 和 p-JNK 水平的特定升高，且呈剂量和时间依赖性。 AGE 给药显着增加了 LOX 和 ET-1 同源启动子区域的 NF-kappa I' 和 AP-1 结合活性。此外，高 AGE 含量饮食后大鼠主动脉内皮中 LOX 和 ET-1 过度表达证实了这些分子的功能相互关系。我们的研究结果表明，AGEs 通过人内皮细胞中的 AGE/RAGE/MAPK 信号级联触发 NF-kappa I' 和 AP-1 介导的 LOX 和 ET-1 上调，从而通过损害内皮屏障功能、改变 ECM 生物力学特性和细胞增殖来破坏内皮稳态。
Endothelial dysfunction involves deregulation of the key extracellular matrix (ECM) enzyme lysyl oxidase (LOX) and the vasoconstrictor protein, endothelin-1 (ET-1), whose gene expression can be modulated by the transcriptional activators nuclear factor kappa B (NF-kappa B) and activator protein-1 (AP-1). Advanced glycation end products (AGEs) present an aggravating factor of endothelial dysfunction which upon engagement to their receptor RAGE induce upregulation of mitogen-activated protein kinases (MAPKs), leading to NF-kappa B and AP-1 potentiation. We hypothesized that AGEs could induce NF-kappa I'- and AP-1-dependent regulation of LOX and ET-1 expression via the AGE/RAGE/MAPK signaling axis. Western blot, real-time qRT-PCR, FACS analysis and electrophoretic mobility-shift assays were employed in human aortic endothelial cells (HAECs) following treatment with AGE-bovine serum albumin (AGE-BSA) to investigate the signaling pathway towards this hypothesis. Furthermore, immunohistochemical analysis of AGEs, RAGE, LOX and ET-1 expression was conducted in aortic endothelium of a rat experimental model exposed to high- or low-AGE content diet. HAECs exposed to AGE-BSA for various time points exhibited upregulation of LOX and ET-1 mRNA levels in a dose- and time-dependent manner. Exposure of HAECs to AGE-BSA also showed specific elevation of phospho(p)-ERK1/2 and p-JNK levels in a dose- and time-dependent fashion. AGE administration significantly increased NF-kappa I'- and AP-1-binding activity to both LOX and ET-1 cognate promoter regions. Moreover, LOX and ET-1 overexpression in rat aortic endothelium upon high-AGE content diet confirmed the functional interrelation of these molecules. Our findings demonstrate that AGEs trigger NF-kappa I'- and AP-1-mediated upregulation of LOX and ET-1 via the AGE/RAGE/MAPK signaling cascade in human endothelial cells, thus contributing to distorted endothelial homeostasis by impairing endothelial barrier function, altering ECM biomechanical properties and cell proliferation.