ATL‐1, an analogue of aspirin‐triggered lipoxin A 4 , is a potent inhibitor of several steps in angiogenesis induced by vascular endothelial growth factor

• Published in: British journal of pharmacology Vol. 153; no. 5; pp. 956 - 965
• Main Authors: Cezar‐de‐Mello, P F T, Vieira, A M, Nascimento‐Silva, V, Villela, C G, Barja‐Fidalgo, C, Fierro, I M
• Format: Journal Article
• Language: English
• Published: 29.01.2009
• ISSN: 0007-1188; 1476-5381
• DOI: 10.1038/sj.bjp.0707650

Background and purpose: Vascular endothelial growth factor (VEGF) is the most important proangiogenic protein. We have demonstrated that ATL‐1, a synthetic analogue of aspirin‐triggered lipoxin A 4 , inhibits VEGF‐induced endothelial cell (EC) migration. In the present study, we investigated the effects of ATL‐1 in several other actions stimulated by VEGF. Methods: Human umbilical vein ECs were treated with ATL‐1 for 30 min before stimulation with VEGF. Cell proliferation was measured by thymidine incorporation. Adherent cells were determined by fluorescence intensity using a Multilabel counter. Expression and activity of matrix metalloproteinases (MMP) were analysed by western blot and zymography. Key results: ATL‐1 inhibited EC adhesion to fibronectin via interaction with its specific receptor. Furthermore, VEGF‐induced MMP‐9 activity and expression were reduced by pretreatment with ATL‐1. Because the transcription factor NF‐κB has been implicated in VEGF‐mediated MMP expression and EC proliferation, we postulated that ATL‐1 might modulate the NF‐κB pathway and, indeed, ATL‐1 inhibited NF‐κB nuclear translocation. Pretreatment of EC with ATL‐1 strongly decreased VEGF‐dependent phosphorylation of phosphainositide 3‐kinase (PI3‐K) and extracellular signal‐regulated kinase‐2 (ERK‐2), two signalling kinases involved in EC proliferation. Inhibition of VEGF‐induced EC proliferation by ATL‐1 was antagonized by sodium orthovanadate, suggesting that this inhibitory activity was mediated by a protein tyrosine phosphatase. This was confirmed by showing that ATL‐1 inhibition of VEGF receptor‐2 (VEGFR‐2) phosphorylation correlates with SHP‐1 association with VEGFR‐2. Conclusions and implications: The synthetic 15‐epi‐lipoxin analogue, ATL‐1, is a highly potent molecule exerting its effects on multiple steps of the VEGF‐induced angiogenesis. British Journal of Pharmacology (2008) 153 , 956–965; doi: 10.1038/sj.bjp.0707650 ; published online 14 January 2008