Abstract 16667: Pro-Inflammatory Effects on Endothelium of High-Density Lipoprotein From Patients With Coronary Artery Disease: Role of Its Proteome Remodeling and PI3k Pathway
Introduction: High-density lipoprotein(HDL) from healthy individual exerts endothelial protection through anti-inflammation. Recent studies revealed that HDL function may shift towards pro-atherogenic when coronary artery disease(CAD) occured.
Hypothesis: HDL from CAD patients promoted endothelium inflammation due to its proteomic remodeling.
Methods: Isolated by ultracentrifugation, plasma HDL samples from CAD patients or healthy volunteers(HDLCAD and HDLhealthy) were each pooled as a different group. Cultured HUVECs were treated with equal amount of HDLCAD or HDLhealthy. Specific PI3K inhibitor Ly294002 was added prior to HDLCAD treatment to investigate pathway involved. MCP-1 and IL-6 levels in the supernatant were measured by ELISA. Differential protein and modification profiles between CAD patients and healthy controls were compared by Isobaric Tags For Relative And Absolute Quantitation(iTRAQ) technology combined with nanoLC-MS/MS method. Gene ontology(GO) category enrichments were applied to analyze the functions of these differential proteins.
Results: HDLCAD stimulated more MCP-1(6.19±1.39 ng/ml vs.0.37±0.09ng/ml,P<0.05) and IL-6 secretion(4.68±0.26ng/ml vs.0.07±0.02ng/ml,P<0.05). Ly294002 decreased the production of both cytokines(MCP-1:2.88±0.89ng/ml vs. 6.19±1.39ng/ml, P<0.05;IL-6:0.18±0.02ng/ml vs. 4.68±0.26ng/ml, P<0.05). Of the 270 proteins identified, 17 were up-regulated, and 6 were down-regulated. 66 proteins in HDL with oxidation, formylation, nitration and phosphorylation were identified. Compared with HDLhealthy, HDLCAD had more oxidized proteins and less nitrationized or phosphorylated proteins(P<0.05).GO analysis showed that all of these proteins were related to lipid metabolism, inflammatory reactions and oxidative stress.
Conclusions: HDLCAD promoted endothelial inflammation through a PI3K-mediated pathway and was associated with the alterations in HDL compostions.
Author Disclosures: Z. Hui: None. L. Hong: None. Y.L. Rong: None. H. Lu: None. L.Y. Shi: None.
- © 2014 by American Heart Association, Inc.