Lysophosphatidylcholine impairs endothelial barrier function through the G protein-coupled receptor GPR4.
Abundant evidence indicates that lysophosphatidylcholine (LPC) is proinflammatory and atherogenic. In the vascular endothelium, LPC increases permeability and expression of proinflammatory molecules such as adhesion molecules and cytokines. Yet, mechanisms by which LPC mediates these activities remain unclear and controversial. Recent evidence implicates involvement of a novel subfamily of G protein-coupled receptors (GPR4, G2A, OGR1, and TDAG8) that are sensitive to lysolipids and protons. We previously reported that one of these receptors, GPR4, is selectively expressed by a variety of endothelial cells and therefore hypothesize that the LPC-stimulated endothelial barrier dysfunction is mediated through GPR4. We developed a peptide Ab against GPR4 that detected GPR4 expression in transfected COS 7 cells and endogenous GPR4 expression in endothelial cells by Western blot. Endothelial cells infected with a retrovirus containing small interference RNA (siRNA) to GPR4 resulted in 40-50% decreased GPR4 expression, which corresponded with partial prevention of the LPC-induced 1) decrease in transendothelial resistance, 2) stress fiber formation, and 3) activation of RhoA. Furthermore, coexpression of the siRNA-GPR4 with a siRNA-resistant mutant GPR4 fully restored the LPC-induced resistance decrease. However, extracellular pH of <7.4 did not alter baseline or LPC-stimulated resistances. The results provide strong evidence that the LPC-mediated endothelial barrier dysfunction is regulated by endogenous GPR4 in endothelial cells and suggest that GPR4 may play a critical role in the inflammatory responses activated by LPC.
Qiao
Jing
J
Rush Univ. Medical Center, Dept. of Pharmacology, 1735 W. Harrison St., Chicago, IL 60612, USA.
Huang
Fei
F
Naikawadi
Ram P
RP
Kim
Kwang S
KS
Said
Tamer
T
Lum
Hazel
H
D000818:Animals; D019556:COS Cells; D002199:Capillary Permeability / Q000187:drug effects* / Q000276:immunology; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D020405:Dermis / Q000098:blood supply; D004553:Electric Conductivity; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D008244:Lysophosphatidylcholines / Q000494:pharmacology*; D016296:Mutagenesis; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism*
C091670:GPR4 protein, human; D008244:Lysophosphatidylcholines; D043562:Receptors, G-Protein-Coupled