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-arachidonoylsiz-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-unsaturatcd 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 chromatolgraphy/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 mug/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 leukocyteendothelial interactions. (C) 2002 Elsevier Science Inc. All rights reserved.