Identification of prostaglandin E2 receptor subtype 2 as a receptor activated by OxPAPC.
Oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine (OxPAPC), which has been shown to accumulate in atherosclerotic lesions and other sites of chronic inflammation, activates endothelial cells (EC) to bind monocytes by activation of endothelial beta1 integrin and subsequent deposition of fibronectin on the apical surface. Our previous studies suggest this function of OxPAPC is mediated via a Gs protein-coupled receptor (GPCR). PEIPC (1-palmitoyl-2-epoxyisoprostane E2-sn-glycero-3-phosphorylcholine) is the most active lipid in OxPAPC that activates this pathway. We screened a number of candidate GPCRs for their interaction with OxPAPC and PEIPC, using a reporter gene assay; we identified prostaglandin E2 receptor EP2 and prostaglandin D2 receptor DP as responsive to OxPAPC. We focused on EP2, which is expressed in ECs, monocytes, and macrophages. OxPAPC component PEIPC, but not POVPC, activated EP2 with an EC50 of 108.6 nmol/L. OxPAPC and PEIPC were also able to compete with PGE2 for binding to EP2 in a ligand-binding assay. The EP2 specific agonist butaprost was shown to mimic the effect of OxPAPC on the activation of beta1 integrin and the stimulation of monocyte binding to endothelial cells. Butaprost also mimicked the effect of OxPAPC on the regulation of tumor necrosis factor-alpha and interleukin-10 in monocyte-derived cells. EP2 antagonist AH6809 blocked the activation of EP2 by OxPAPC in HEK293 cells and blocked the interleukin-10 response to PEIPC in monocytic THP-1 cells. These results suggest that EP2 functions as a receptor for OxPAPC and PEIPC, either as the phospholipid ester or the released fatty acid, in both endothelial cells and macrophages.
Li
Rongsong
R
Department of Pathology, University of California, Los Angeles, CA 90095, USA.
Mouillesseaux
Kevin P
KP
Montoya
Dennis
D
Cruz
Daniel
D
Gharavi
Navid
N
Dun
Martin
M
Koroniak
Lukasz
L
Berliner
Judith A
JA
D000527:Alprostadil / Q000031:analogs & derivatives / Q000494:pharmacology; D050197:Atherosclerosis / Q000209:etiology*; D002478:Cells, Cultured; D015232:Dinoprostone / Q000378:metabolism; D005487:Foam Cells / Q000502:physiology; D006801:Humans; D016753:Interleukin-10 / Q000096:biosynthesis; D028421:Isoprostanes / Q000378:metabolism / Q000494:pharmacology; D008264:Macrophages / Q000378:metabolism; D009000:Monocytes / Q000378:metabolism; D010084:Oxidation-Reduction; D010713:Phosphatidylcholines / Q000378:metabolism / Q000494:pharmacology; D010742:Phospholipid Ethers / Q000378:metabolism / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis; D018078:Receptors, Prostaglandin E / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D058307:Receptors, Prostaglandin E, EP2 Subtype; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis; D044004:Xanthones / Q000494:pharmacology
C400184:1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphorylcholine; C500237:1-palmitoyl-2-(epoxyisoprostane-E2)-sn-glycero-3-phosphocholine; D028421:Isoprostanes; C545644:PTGER2 protein, human; D010713:Phosphatidylcholines; D010742:Phospholipid Ethers; D012333:RNA, Messenger; D018078:Receptors, Prostaglandin E; D058307:Receptors, Prostaglandin E, EP2 Subtype; D014409:Tumor Necrosis Factor-alpha; D044004:Xanthones; D016753:Interleukin-10; C053876:6-isopropoxy-9-oxoxanthene-2-carboxylic acid; D000527:Alprostadil; C048491:butaprost; D015232:Dinoprostone