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ADP Stimulates Human Endothelial Cell Migration via P2Y1 Receptor‐mediated MAPK Pathways
Author(s) -
Shen Jianzhong,
DiCorleto Paul E.
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.a1162-c
Subject(s) - mapk/erk pathway , microbiology and biotechnology , phosphorylation , p2y receptor , kinase , p2y12 , umbilical vein , p38 mitogen activated protein kinases , human umbilical vein endothelial cell , receptor , purinergic signalling , signal transduction , adenosine , platelet activation , endothelial stem cell , biology , chemistry , purinergic receptor , pharmacology , adenosine receptor , endocrinology , biochemistry , platelet , agonist , immunology , in vitro , platelet aggregation
Extensive research on the role of ADP in platelet activation led to the design of new anti‐thrombotic drugs, such as Plavix; however, very little is known about the ADP‐preferring nucleotide receptors (P2Y1, P2Y12, P2Y13) in endothelium. Here, we show that ADP stimulated migration of cultured human umbilical vein endothelial cells (HUVEC) in both Boyden chamber and in vitro wound repair assays. This pro‐migratory effect was mimicked by 2‐MsADP, but not by AMP, and was inhibited by MRS2179 (P2Y1 antagonist), but not by AR‐C69931MX (P2Y12/13 antagonist), suggesting a role for the P2Y1 receptor. Consistent with this interpretation, RT‐PCR revealed abundant P2Y1, barely detectable P2Y12, and absent P2Y13 receptor message in these cells. In addition, both ADP and 2‐MsADP, but not AMP and adenosine, activated the MAPK pathways as evidenced by increased phosphorylation of ERK1/2, JNK and P38 kinases. ADP also stimulated phosphorylation of p90RSK, a downstream substrate of phosphorylated ERK1/2, and induced phosphorylation of several transcription factors downstream of the JNK and P38 pathways, including ATF‐2 and c‐JUN. These signaling events were inhibited by MRS2179, but not by AR‐C69931MX. Furthermore, blockade of the P2Y1 receptor (MRS2179), or the MAPK pathways, including ERK (U0126), JNK (SP600125) or P38 (SB203580), abolished or decreased ADP‐ and 2‐MsADP‐induced HUVEC migration. We conclude that ADP promotes human endothelial cell migration by activating P2Y1 receptor‐mediated MAPK pathways, possibly contributing to re‐endothelialization and angiogenesis after vascular injury. Support: NIH HL29582 and RR00080.