ono-ae1-329 and butaprost

The prostaglandin D. In this study, we investigated an involvement of PGE. Expression of EP2 and EP4, but not EP1 and EP3, was decreased in blood eosinophils of patients with EoE vs. control subjects. Adhesion of eosinophils to esophageal epithelial cells was decreased by EP2 receptor agonist butaprost and EP4 agonist ONO-AE1-329, whereas DP1 agonist BW245C increased adhesion. In chemotaxis assays with supernatant from human esophageal epithelial cells, only ONO-AE1-329 but not butaprost or BW245C inhibited the migration of eosinophils. Expression of EP and DP receptors in epithelial cells and eosinophils was detected in sections of esophageal biopsies from EoE patients by immunohistochemistry. qPCR of biopsies from EoE patients revealed that gene expression of EP4 and DP1 was the highest among PGE. Activation of EP2 and EP4 receptors may inhibit eosinophil recruitment to the esophageal mucosa. However, their activation could negatively affect esophageal barrier integrity suggesting that eosinophilic rather than epithelial EP2 and EP4 have a protective role in EoE.

Topics: Alprostadil; Cell Adhesion; Cell Migration Assays; Cells, Cultured; Eosinophilic Esophagitis; Eosinophils; Esophageal Mucosa; Humans; Immunohistochemistry; Methyl Ethers; Pilot Projects; Prostaglandins E, Synthetic; Receptors, Prostaglandin E, EP2 Subtype; Receptors, Prostaglandin E, EP4 Subtype

Prostaglandin E2 (PGE2), a major metabolite of arachidonic acid produced by cyclooxygenase pathways, exerts its bioactive responses by activating four E-prostanoid receptor subtypes, EP1, EP2, EP3, and EP4. PGE2 enables modulating N-methyl-D-aspartate (NMDA) receptor-mediated responses. However, the effect of E-prostanoid receptor agonists on large-conductance Ca(2+)-activated K(+) (BK) channels, which are functionally coupled with NMDA receptors, remains unclear. Here, we showed that EP2 receptor-mediated signaling pathways increased NMDA-induced outward currents (I NMDA-OUT), which are associated with the BK channel activation. Patch-clamp recordings from the acutely dissociated mouse cortical neurons revealed that an EP2 receptor agonist activated I NMDA-OUT, whereas an EP3 receptor agonist reduced it. Agonists of EP1 or EP4 receptors showed no significant effects on I NMDA-OUT. A direct perfusion of 3,5'-cyclic adenosine monophosphate (cAMP) through the patch pipette facilitated I NMDA-OUT, which was abolished by the presence of protein kinase A (PKA) inhibitor. Furthermore, facilitation of I NMDA-OUT caused by an EP2 receptor agonist was significantly suppressed by PKA inhibitor. Finally, the activation of BK channels through EP2 receptors facilitated the recovery phase of NMDA-induced dendritic beading in the primary cultured cortical neurons. These results suggest that a direct activation of BK channels by EP2 receptor-mediated signaling pathways plays neuroprotective roles in cortical neurons.

Topics: Alprostadil; Animals; Carbazoles; Dinoprostone; In Vitro Techniques; Indoles; Large-Conductance Calcium-Activated Potassium Channels; Male; Methyl Ethers; Mice; Mice, Inbred C57BL; Pyrroles; Receptors, N-Methyl-D-Aspartate; Receptors, Prostaglandin E, EP1 Subtype; Receptors, Prostaglandin E, EP2 Subtype; Receptors, Prostaglandin E, EP3 Subtype; Receptors, Prostaglandin E, EP4 Subtype

Prostaglandin E2 (PGE2) is elevated during cardiac injury and we have previously shown that mice lacking the PGE2 EP4 receptor display dilated cardiomyopathy (DCM) with increased expression of the membrane type matrix metalloproteinase, MMP-14. We thus hypothesized that PGE2 regulates expression of MMP-14 and also affects fibroblast migration. Primary cultures of neonatal rat ventricular fibroblasts (NVFs) were used to test the effects of PGE2. Gene and protein expression was assessed by real time RT-PCR and Western blot, MMP activity was determined by zymography and migration of NVF was assessed by motility in a transwell system. PGE2 reduced expression of MMP-14 and these effects were antagonized by an EP4 antagonist. An EP4 agonist mimicked the effect of PGE2. PGE2 also increased mRNA and protein levels of plasminogen activator inhibitor-1 (PAI-1), an inhibitor of MMP activation. However, PGE2-stimulation of PAI-1 was mediated by the EP1/EP3 receptor and not EP4. Migration of NVF was assessed by motility in a transwell system. Treatment of NVFs with PGE2 reduced the number of cells migrating toward 10% FCS. Treatment with the EP2 agonist also reduced migration but did not affect MMP-14 expression or PAI-1. Our results suggest that PGE2 utilizes different receptors and mechanisms to ultimately decrease MMP expression and NVF migration.

Topics: Alprostadil; Animals; Animals, Newborn; Cardiomyopathies; Cell Movement; Dinoprostone; Fibroblasts; Gene Expression Regulation; Male; Matrix Metalloproteinase 14; Methyl Ethers; Naphthalenes; Phenylbutyrates; Plasminogen Activator Inhibitor 1; Rats; Real-Time Polymerase Chain Reaction; Receptors, Prostaglandin E, EP4 Subtype; RNA, Messenger

Mice deficient in the G-protein alpha subunit G(i)alpha(2) spontaneously develop colitis and colon cancer. IL-11 is a pleiotropic cytokine known to protect the intestinal epithelium from injury in animal models of colitis and is produced by subepithelial myofibroblasts in response to inflammatory mediators including TGF-beta, IL-1beta, and PGE(2). Arachidonic acid release and subsequent PGE(2) production is significantly decreased in the colonic mucosa of G(i)alpha(2)-/- mice, and we hypothesized that this would affect mucosal IL-11 production. Mucosal levels of IL-11 were found to be significantly decreased in G(i)alpha(2)-/- mice despite the presence of mild colitis. Primary cultures of G(i)alpha(2)-/- intestinal and colonic myofibroblasts (IMF and CMF, respectively) produced less basal and TGF-beta or IL-1beta-stimulated IL-11 mRNA and protein than wild-type cells. Inhibitors of ERK or p38 MAPK activation dose dependently inhibited IMF and CMF IL-11 production in response to TGF-beta stimulation, whereas 16,16 dimethyl-PGE(2) and prostanoid receptor subtype-selective agonists induced IL-11 production. Treatment of animals with the EP4-specific agonist ONO-AE1-329 resulted in enhanced mucosal levels of IL-11, and increased IL-11 production by ex vivo cultured CMF. Modulation of cAMP levels produced diverging results, with enhancement of TGF-beta-induced IL-11 release in IMF pretreated with 8-Br-cAMP and inhibition in cells treated either with pertussis toxin or the PKA inhibitor H-89. These data suggest a physiological role for prostaglandins, MAPK signaling, and cAMP signaling for the production of myofibroblast-derived IL-11 in the mouse intestinal mucosa.

Topics: 16,16-Dimethylprostaglandin E2; Alprostadil; Animals; Cells, Cultured; Colon; Cyclic AMP; Dinoprostone; Dose-Response Relationship, Drug; Enzyme Activation; Extracellular Signal-Regulated MAP Kinases; Fibroblasts; Flavonoids; GTP-Binding Protein alpha Subunit, Gi2; Imidazoles; Interleukin-11; Interleukin-1beta; Intestine, Small; Methyl Ethers; Mice; Mice, Inbred C57BL; Mice, Knockout; Mitogen-Activated Protein Kinases; p38 Mitogen-Activated Protein Kinases; Protein Kinase Inhibitors; Pyridines; Receptors, Prostaglandin E; Receptors, Prostaglandin E, EP4 Subtype; RNA, Messenger; Signal Transduction; Transforming Growth Factor beta

Prostaglandin E2 (PGE2) exerts its biological actions via EP receptors (EP1, EP2, EP3, and EP4). In the present study, we investigated whether PGE2 regulated interleukin (IL)-1beta-induced matrix metalloproteinase (MMP)-3 production in human gingival fibroblasts (HGF) derived from periodontally healthy subjects and diseased patients. In HGF from healthy gingiva, PGE2 down-regulated IL-1beta-induced MMP-3 production, whereas in HGF from periodontitis patients, PGE2 enhanced it. Butaprost (an EP2 agonist) and ONO-AE1-329 (an EP4 agonist) suppressed IL-1beta-induced MMP-3 production, and 17-phenyl-omega-trinor PGE2 (an EP1 agonist) mimicked the PGE(2) effect in HGF from healthy and periodontally diseased tissues, respectively. Analysis of these data suggests that, in HGF from healthy tissue, IL-1beta-induced MMP-3 production is down-regulated by PGE2 via EP2 and EP4 receptors, whereas in cells from periodontally diseased tissue, IL-1beta-induced MMP-3 production is up-regulated via EP1 receptors. Different regulation of IL-1beta-induced MMP-3 production by PGE2 between healthy and periodontally diseased tissues may be involved in the pathogenesis of periodontal disease.

Topics: Adult; Alprostadil; Cells, Cultured; Dinoprostone; Down-Regulation; Fibroblasts; Gingiva; Humans; Interleukin-1; Matrix Metalloproteinase 3; Matrix Metalloproteinase Inhibitors; Methyl Ethers; Middle Aged; Periodontitis; Prostaglandins E, Synthetic; Receptors, Prostaglandin E; Receptors, Prostaglandin E, EP1 Subtype; Receptors, Prostaglandin E, EP4 Subtype; Up-Regulation

Although previous pharmacologic studies have indicated that PGE receptors are expressed in human eosinophils, the exact distribution of the subtypes remains mostly unknown. By using a combination of genetic and conventional pharmacologic approaches, coexpression of mRNAs encoding the PGE receptor 2 (EP2) and EP4 was confirmed in eosinophils. Moreover, competitive PCR analysis of eosinophil RNA revealed that levels of the EP4 receptor mRNA were significantly higher than those of the EP2 receptor mRNA (P =.04). On the basis of the expression levels of mRNAs, an EP4 agonist, but not an EP2 agonist, was effective in inducing cyclic AMP production in eosinophils, suggesting that the EP4 receptor is of primary importance in eosinophil functions of PGE(2).

Topics: Alprostadil; Cyclic AMP; Dinoprostone; Dose-Response Relationship, Drug; Eosinophils; Humans; Methyl Ethers; Receptors, Prostaglandin E; Receptors, Prostaglandin E, EP2 Subtype; Receptors, Prostaglandin E, EP4 Subtype; RNA, Messenger