Involvement of cAMP-response element binding protein-1 in arachidonic acid-induced vascular smooth muscle cell motility

Abstract

In addition to their role in many vital cellular functions, arachidonic acid (AA) and its eicosanoid metabolites are involved in the pathogenesis of several diseases, including atherosclerosis and cancer. To understand the potential mechanisms by which these lipid molecules could influence the disease processes, particularly cardiovascular diseases, we studied AA's effects on vascular smooth muscle cell (VSMC) motility and the role of cAMPresponse element binding protein-1 (CREB-1) in this process. AA exerted differential effects on VSMC motility; at lower doses, it stimulated motility, whereas at higher doses, it was inhibitory. AA-induced VSMC motility requires its conversion via the lipoxygenase (LOX) and cyclooxygenase (COX) pathways. AA stimulated the phosphorylation of extracellular signal-regulated kinases (ERKs), Jun N-terminal kinases ( JNKs), and p38 mitogen-activated protein kinase (p38MAPK) in a time-dependent manner, and blockade of these serine/threonine kinases significantly attenuated AAinduced VSMC motility. In addition, AA stimulated CREB-1 phosphorylation and activity in a manner that was also dependent on its metabolic conversion via the LOX and COX pathways and the activation of ERKs and p38MAPK but not JNKs. Furthermore, suppression of CREB-1 activation inhibited AA-induced VSMC motility. 15(S)-Hydroxyeicosatetraenoic acid and prostaglandin F 2a , the 15-LOX and COX metabolites of AA, respectively, that are produced by VSMC at lower doses, were also found to stimulate motility in these cells. Together, these results suggest that AA induces VSMC motility by complex mechanisms involving its metabolism via the LOX and COX pathways as well as the ERK-and p38MAPK-dependent and JNK-independent activation of CREB-1.-Dronadula, N., F. Rizvi, E. Blaskova, Q. Li, and G. N. Rao. Involvement of cAMP-response element binding protein-1 in arachidonic acid-induced vascular smooth muscle cell motility. J. Lipid Res. 2006. 47: 767-777.

Nuclear Factor of Activated T Cells c1 Mediates p21-activated Kinase 1 Activation in the Modulation of Chemokine-induced Human Aortic Smooth Muscle Cell F-actin Stress Fiber Formation, Migration, and Proliferation and Injury-induced Vascular Wall Remodeling

“…Cell Migration-Cell migration was measured using a modified Boyden chamber method as described previously (28). Cell motility is presented as number of migrated cells/field.…”

Identification of a cAMP-response Element in the Regulator of G-protein Signaling-2 (RGS2) Promoter as a Key Cis-regulatory Element for RGS2 Transcriptional Regulation by Angiotensin II in Cultured Vascular Smooth Muscles

“…Electrophoretic Mobility Shift Assay (EMSA)-Nuclear extracts were prepared and EMSA was performed as described (31,32). Briefly, nuclear extracts (2 g of protein) were incubated (30 min on ice) with 30-mer 32 P-labeled double-stranded WT or mutant RGS2-CRE1 probes in a mixture (15 l total volume) consisting of 15 mM HEPES, pH 7.9, 3 mM Tris-HCl, pH 7.9, 60 mM KCl, 1 mM EDTA, 1 mM PMSF, 1 mM dithiothreitol, 4.5 g of BSA, 2 g of poly(dI-dC), and 15% glycerol.…”

“…4, A and B, contain CREB, three independent approaches were employed. First, a consensus CRE oligonucleotide that is known to bind to CREB was used as a competitor (31). The appearance of shifted bands induced by FSK (Fig.…”

15-Lipoxygenase-1-enhanced Src-Janus Kinase 2-Signal Transducer and Activator of Transcription 3 Stimulation and Monocyte Chemoattractant Protein-1 Expression Require Redox-sensitive Activation of Epidermal Growth Factor Receptor in Vascular Wall Remodeling

“…Cell Migration-Cell migration was measured using a modified Boyden chamber method as described previously (29). Wherever adenovirus was used, cells were infected with the respective adenovirus at an m.o.i.…”