Abstract
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-κB) 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-κB and AP-1 potentiation. We hypothesized that AGEs could induce NF-κΒ- 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-κΒ- 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-κΒ- 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.
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Abbreviations
- AGEs:
-
Advanced glycation end products
- AP-1:
-
Activator protein-1
- BAECs:
-
Bovine aortic endothelial cells
- COX-2:
-
Cyclo-oxygenase-2
- CRP:
-
C-reactive protein
- CVD:
-
Cardiovascular disease
- ET-1:
-
Endothelin-1
- HDL:
-
High-density lipoprotein
- ICAM-1:
-
Intercellular adhesion molecule-1
- IL-1β:
-
Interleukin-1 beta
- LDL:
-
Low-density lipoprotein
- LOX:
-
Lysyl oxidase
- MAPK:
-
Mitogen-activated protein kinase
- NF-κB:
-
Nuclear factor kappa B
- NO:
-
Nitric oxide
- PCOS:
-
Polycystic ovary syndrome
- RAGE:
-
Receptor for AGEs
- ROS:
-
Reactive oxygen species
- VEGF:
-
Vascular endothelial growth factor
- VSMCs:
-
Vascular smooth muscle cells
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Adamopoulos, C., Piperi, C., Gargalionis, A.N. et al. 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. Cell. Mol. Life Sci. 73, 1685–1698 (2016). https://doi.org/10.1007/s00018-015-2091-z
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DOI: https://doi.org/10.1007/s00018-015-2091-z