Hydroxy alkenal phospholipids regulate inflammatory functions of endothelial cells.
Monocyte recruitment into the vessel wall plays an important role in atherogenesis. Polar lipid components of minimally modified/oxidized LDL were shown to activate endothelial cells to increase the synthesis of monocyte chemotactic factors and surface expression of adhesion molecules. We previously reported regulation of endothelial cell inflammatory functions by oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (Ox-PAPC) and three component oxidized phospholipids, containing oxovaleroyl (POVPC), glutaroyl (PGPC) and epoxyisoprostane (PEIPC) groups at the sn-2 position of oxidized phospholipids. In the present study, we demonstrate the presence of gamma-hydroxy-alpha,beta-unsaturated aldehydic phospholipid, 1-palmitoyl-2-(5-hydroxy-8-oxooct-6-enoyl)-sn-glycero-3-phosphocholine (HOOA-PC; m/z 650.4), in Ox-PAPC by liquid chromatography/mass spectrometry (LC/MS), LC/MS/MS, derivatization and tandem mass spectrometric analyses. This was further unambiguously confirmed by the identical chromatographic and mass spectrometric characteristics of Ox-PAPC-derived m/z 650.4 with synthetic HOOA-PC. The time course of PAPC autoxidation showed that HOOA-PC accumulates with oxidation and represents about 2% of Ox-PAPC. We have also examined the effects of HOOA-PC on leukocyte-endothelial interactions. HOOA-PC dose-dependently activated human aortic endothelial cells (HAECs) to bind monocytes (twofold at 10 micrograms/ml) and caused a dose-dependent increase (two- to threefold) in levels of monocyte chemotactic protein-1 (MCP-1) and interleukin-8 (IL-8)--chemokines that are important in monocyte entry into chronic lesions. HOOA-PC also inhibited LPS-induced expression of E-Selectin, a major adhesion molecule that mediates neutrophil endothelial interactions. The present study suggests that the HOOA-PC exerts its effects on endothelial cells as a free lipid. These studies demonstrate the importance of HOOA-PC as a new potential proinflammatory molecule that regulates leukocyte-endothelial interactions.
Subbanagounder
Ganesamoorthy
G
Department of Medicine/Cardiology, Center for the Health Sciences, Room 47-123, UCLA Medical Center, Los Angeles, CA 90095-1679, USA. gsubbanagounder@mednet.ucla.edu
Deng
Yujin
Y
Borromeo
Christine
C
Dooley
Alek N
AN
Berliner
Judith A
JA
Salomon
Robert G
RG
D000893:Anti-Inflammatory Agents / Q000737:chemistry / Q000494:pharmacology*; D002478:Cells, Cultured; D018925:Chemokines / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D019040:E-Selectin / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000473:pathology*; D004797:Enzyme-Linked Immunosorbent Assay; D005997:Glycerylphosphorylcholine / Q000378:metabolism; D006801:Humans; D007249:Inflammation / Q000473:pathology*; D008070:Lipopolysaccharides / Q000494:pharmacology; D013058:Mass Spectrometry; D009829:Oleic Acids / Q000494:pharmacology; D010084:Oxidation-Reduction; D010742:Phospholipid Ethers / Q000737:chemistry / Q000378:metabolism; D010743:Phospholipids / Q000494:pharmacology*; D021241:Spectrometry, Mass, Electrospray Ionization; D013997:Time Factors
C400184:1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphorylcholine; C102523:6-hydroxyoleic acid; D000893:Anti-Inflammatory Agents; D018925:Chemokines; D019040:E-Selectin; D008070:Lipopolysaccharides; D009829:Oleic Acids; D010742:Phospholipid Ethers; D010743:Phospholipids; D005997:Glycerylphosphorylcholine