%0 Journal Article %A Alatangaole Damirin %A Hideaki Tomura %A Mayumi Komachi %A Masayuki Tobo %A Koichi Sato %A Chihiro Mogi %A Hiromi Nochi %A Koichi Tamoto %A Fumikazu Okajima %T Sphingosine 1-Phosphate Receptors Mediate the Lipid-Induced cAMP Accumulation through Cyclooxygenase-2/Prostaglandin I2 Pathway in Human Coronary Artery Smooth Muscle Cells %D 2005 %R 10.1124/mol.104.004317 %J Molecular Pharmacology %P 1177-1185 %V 67 %N 4 %X Sphingosine 1-phosphate (S1P) has been shown to exert a variety of biological responses through extracellular specific receptors or intracellular mechanisms. In the present study, we characterized a signaling pathway of S1P-induced cAMP accumulation in human coronary artery smooth muscle cells (CASMCs). S1P induced biphasic cAMP accumulation composed of a short-term and transient response (a peak at 2.5 min) and a late and sustained response (∼4-6 h). The late phase of cAMP accumulation was parallel to the increment of cyclooxygenase-2 protein expression and was inhibited by N-[2-(cyclohexyloxyl)-4-nitrophenyl]-methane sulfonamide (NS398), a cyclooxygenase-2-specific inhibitor. We were surprised to find that the cyclooxygenase-2 inhibitor also inhibited short-term cAMP accumulation even when cyclooxygenase-2 protein expression was not yet increased. More interestingly, the short-term cAMP accumulation was also completely inhibited by pertussis toxin, an inhibitor of Gi/o proteins. JTE-013, a specific antagonist for S1P2 receptors, inhibited the S1P-induced cAMP accumulation. Furthermore, small interfering RNAs targeted for S1P2 receptors significantly inhibited the S1P-induced cAMP accumulation. The cAMP response was also inhibited by specific inhibitors for phospholipase C, extracellular signal-regulated kinase pathways, and cytosolic phospholipase A2. S1P actually activated these enzyme activities and stimulated prostaglandin I2 (PGI2) synthesis. Finally, exogenously applied arachidonic acid and PGI2 induced cAMP accumulation to a similar extent as S1P. In conclusion, S1P induced cAMP accumulation through S1P receptors, including S1P2 receptor and Gi/o protein-mediated stimulation of intracellular signaling pathways involving cyclooxygenase-2-dependent PGI2 synthesis. %U https://molpharm.aspetjournals.org/content/molpharm/67/4/1177.full.pdf