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(Circulation Research. 2008;102:448.)
© 2008 American Heart Association, Inc.

Molecular Medicine

ADP Stimulates Human Endothelial Cell Migration via P2Y₁ Nucleotide Receptor–Mediated Mitogen-Activated Protein Kinase Pathways

Jianzhong Shen, Paul E. DiCorleto

From the Department of Cell Biology, The Lerner Research Institute, Cleveland Clinic Foundation, Ohio.

Correspondence to Paul E. DiCorleto, PhD, Professor and Chairman, The Lerner Research Institute, Cleveland Clinic Lerner College of Medicine, Cleveland Clinic Foundation, 9500 Euclid Ave, Cleveland, OH 44195. E-mail dicorlp{at}ccf.org

Extensive research on the role of ADP in platelet activation led to the design of new anti-thrombotic drugs, such as clopidogrel (Plavix; sanofi-aventis); however, very little is known about the ADP-preferring nucleotide receptors (P2Y₁, P2Y₁₂, and P2Y₁₃) in endothelium. Here, we show that ADP stimulates migration of cultured human umbilical vein endothelial cells (HUVECs) in both Boyden chamber and in vitro wound repair assays. This promigratory effect was mimicked by 2-MeSADP, but not by AMP, and was inhibited by MRS2179 (P2Y₁ receptor antagonist) but not by AR-C69931MX (P2Y_12/13 receptor antagonist). RT-PCR revealed abundant P2Y₁, barely detectable P2Y₁₂, and absent P2Y₁₃ receptor message in these cells. In addition, both ADP and 2-MeSADP, but not AMP, activated the mitogen-activated protein kinase pathways as evidenced by increased phosphorylation of extracellular signal-regulated kinase (ERK)1/2, c-Jun N-terminal kinase (JNK), and p38 kinase. ADP also stimulated phosphorylation of p90RSK, a downstream substrate of phosphorylated ERK1/2, and induced phosphorylation of such transcription factors downstream of the JNK and p38 pathways as c-Jun and activating transcription factor-2. These signaling events were inhibited by MRS2179 but not by AR-C69931MX. Furthermore, blockade of the ERK or JNK pathways by U0126 and SP600125, respectively, abolished ADP- and 2-MeSADP–stimulated HUVEC migration. However, inhibition of the p38 pathway by SB203580 partially suppressed ADP- and 2-MeSADP–induced HUVEC migration. We conclude that ADP promotes human endothelial cell migration by activating P2Y₁ receptor–mediated MAPK pathways, possibly contributing to reendothelialization and angiogenesis after vascular injury.

Key Words: nucleotide receptors • vascular endothelial cells • protein kinases • cell migration

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