Abstract 657: Resolvin D1 Attenuates Vascular Smooth Muscle Cell Migration via the cAMP Pathway
Introduction: Resolvin D1 (RvD1), a specialized pro-resolving lipid mediator (SPM), attenuates the migratory phenotype in vascular smooth muscle cells (VSMC), which is critical to the development of atherosclerosis and neointimal hyperplasia. SPM are known to interact with G-protein coupled receptors (GPCR). We sought to investigate the biochemical pathways by which RvD1 influences VSMC migration.
Methods: VSMC were harvested from human saphenous veins. cAMP levels were measured via ELISA, in the absence or presence of WRW4, a blocking peptide for the RvD1 receptor ALX, a GPCR. PDGF-BB (10ng/ml), thrombin (1U/ml) and angiotensin (AT II; 1μM) were used as agonists in a VSMC scratch assay. PDGF-BB-induced cytoskeletal changes were measured as length to width ratio after actin-phalloidin staining. Rp-8-Br-cAMP (10μM), a selective PKA inhibitor, was used in the phenotypic assays.
Results: RvD1 treatment (10nM) of VSMC induced a significant acute flux in cAMP levels, causing a dramatic increase at 5 minutes (Fig. 1A; n≥3); this increase was abolished by WRW4. RvD1 (10nM) attenuated VSMC migration stimulated by PDGF-BB, thrombin and AT II (Fig. 1B; n≥3) and cytoskeletal rearrangements stimulated by PDGF-BB (n=4). RvD1’s effects were negated by the addition of Rp-8-Br-cAMP, suggesting a central role for the cAMP pathway.
Conclusion: Our results suggest that RvD1 attenuates VSMC migration by increasing levels of cAMP through ALX.
Author Disclosures: G. Mottola: None. B. Wu: None. A. Chatterjee: None. M. Chen: None. S.R. Komshian: None. M.S. Conte: None.
- © 2015 by American Heart Association, Inc.