Vascular and inflammatory stresses mediate atherosclerosis via RAGE and its ligands in apoE–/– mice
Evis Harja, De-xiu Bu, Barry I. Hudson, Jong Sun Chang, Xiaoping Shen, Kellie Hallam, Anastasia Z. Kalea, Yan Lu, Rosa H. Rosario, Sai Oruganti, Zana Nikolla, Dmitri Belov, Evanthia Lalla, Ravichandran Ramasamy, Shi Fang Yan, Ann Marie Schmidt
Evis Harja, De-xiu Bu, Barry I. Hudson, Jong Sun Chang, Xiaoping Shen, Kellie Hallam, Anastasia Z. Kalea, Yan Lu, Rosa H. Rosario, Sai Oruganti, Zana Nikolla, Dmitri Belov, Evanthia Lalla, Ravichandran Ramasamy, Shi Fang Yan, Ann Marie Schmidt
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Research Article Cardiology

Vascular and inflammatory stresses mediate atherosclerosis via RAGE and its ligands in apoE–/– mice

Abstract

Endothelial dysfunction is a key triggering event in atherosclerosis. Following the entry of lipoproteins into the vessel wall, their rapid modification results in the generation of advanced glycation endproduct epitopes and subsequent infiltration of inflammatory cells. These inflammatory cells release receptor for advanced glycation endproduct (RAGE) ligands, specifically S100/calgranulins and high-mobility group box 1, which sustain vascular injury. Here, we demonstrate critical roles for RAGE and its ligands in vascular inflammation, endothelial dysfunction, and atherosclerotic plaque development in a mouse model of atherosclerosis, apoE–/– mice. Experiments in primary aortic endothelial cells isolated from mice and in cultured human aortic endothelial cells revealed the central role of JNK signaling in transducing the impact of RAGE ligands on inflammation. These data highlight unifying mechanisms whereby endothelial RAGE and its ligands mediate vascular and inflammatory stresses that culminate in atherosclerosis in the vulnerable vessel wall.

Authors

Evis Harja, De-xiu Bu, Barry I. Hudson, Jong Sun Chang, Xiaoping Shen, Kellie Hallam, Anastasia Z. Kalea, Yan Lu, Rosa H. Rosario, Sai Oruganti, Zana Nikolla, Dmitri Belov, Evanthia Lalla, Ravichandran Ramasamy, Shi Fang Yan, Ann Marie Schmidt

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Figure 5

RAGE-mediated upregulation of inflammatory molecules in murine aortic endothelial cells: oxLDL.

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RAGE-mediated upregulation of inflammatory molecules in murine aortic en...
(A) oxLDL contains AGE epitopes. Native LDL and oxLDL (5 μg/ml) were subjected to SDS-PAGE and Western blotting using affinity-purified rabbit anti-AGE IgG. (B) Endothelial cells were incubated with 5 μg/ml oxLDL for 4 hours. Western blotting was performed for detection of VCAM-1 antigen followed by anti–β-actin IgG. (C) Zymography for detection of MMP-2 activity was performed. *P < 0.0001 versus unstimulated WT; **P < 0.0001 versus stimulated WT. (D and E) Effect of anti-AGE IgG. Murine aortic endothelial cells were pretreated with rabbit anti-AGE IgG or nonimmune rabbit IgG (nI-IgG; 50 μg/ml) for 1 hour. OxLDL (5 μg/ml) was added for 4 hours; cells were harvested and Western blotting was performed to detect VCAM-1 (D) followed by anti–β-actin IgG, or MMP-2 activity by zymography (E). (F and G) Signal transduction. Endothelial cells were incubated with 5 μg/ml oxLDL for 20 minutes. Western blotting for detection of phospho/total pERK and JNK MAP kinases was performed. (H) Endothelial cells were pretreated with the pERK MAP kinase inhibitor PD98059 (10 μM) or the JNK MAP kinase inhibitor SP600125 (10 μM) for 1 hour prior to exposure to oxLDL for 20 minutes. Western blotting was performed for detection of VCAM-1 antigen. *P < 0.0001 versus unstimulated WT; **P < 0.0001 versus oxLDL-stimulated WT.