thromboxane-a2 and cicaprost

1. The prostanoid receptor(s) that mediates inhibition of bacterial lipopolysaccharide (LPS)-induced tumour necrosis factor-alpha (TNF-alpha) generation from human peripheral blood monocytes was classified by use of naturally occurring and synthetic prostanoid agonists and antagonists. 2. In human monocytes that were adherent to plastic, neither prostaglandin D2 (PGD2), prostaglandin E2 (PGE2), prostaglandin F(2 alpha) (PGF(2 alpha)) nor the stable prostacyclin and thromboxane mimetics, cicaprost and U-46619, respectively, promoted the elaboration of TNF alpha-like immunoreactivity, as assessed with a specific ELISA, indicating the absence of excitatory prostanoid receptors on these cells. 3. Exposure of human monocytes to LPS (3 ng ml-1, approximately EC84) resulted in a time-dependent elaboration to TNF alpha which was suppressed in cells pretreated with prostaglandin E1 (PGe1), PGE2 and cicaprost. This effect was concentration-dependent with mean pIC50 values of 7.14, 7.34 and 8.00 for PGE1, PGE2 and cicaprost, respectively. PGD2, PGF(2 alpha) and U-46619 failed to inhibit the generation of TNF alpha at concentrations up to 10 microM. 4. With respect to PGE2, the EP-receptor agonists, 16,16-dimethyl PGE2 (non-selective), misoprostol (EP2/EP3-selective), 11-deoxy PGE1 (EP2-selective) and butaprost (EP2-selective) were essentially full agonists as inhibitors of LPS-induced TNF alpha generation with mean pIC50 values of 6.21, 6.02, 5.67 and 5.59, respectively. In contrast to the results obtained with butaprost and 11-deoxy PGE1, another EP2-selective agonist, AH 13205, inhibited TNF alpha generation by only 21% at the highest concentration (10 microM) examined. EP-receptor agonists which have selectively for the EP1- (17-phenyl-omega-trinor PGE2) and EP3-receptor (MB 28,767, sulprostone) were inactive or only weakly active as inhibitors of TNF alpha generation. 5. Pretreatment of human monocytes with the TP/EP4-receptor antagonist, AH 23848B, at 10, 30 and 100 microM suppressed LPS-induced TNF alpha generation by 10%, 28% and 77%, respectively, but failed to shift significantly the location of the PGE2 concentration-response curves. 6. Given that AH 13205 was a poor inhibitor of TNF alpha generation, studies were performed to determine if it was a partial agonist and whether it could antagonize the inhibitory effect of PGE2. Pretreatment of human monocytes with 10 and 30 microM AH 13205 inhibited the generation of TNF alpha by 31% and 53%, respectively, b

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Biphenyl Compounds; Dinoprostone; Drug Interactions; Epoprostenol; Humans; Leukocytes, Mononuclear; Lipopolysaccharides; Prostaglandin Antagonists; Prostaglandin Endoperoxides, Synthetic; Prostanoic Acids; Receptors, Prostaglandin; Thromboxane A2; Tumor Necrosis Factor-alpha; Xanthenes; Xanthones

1. The influence of endogenously produced prostanoids on active transepithelial Ca2+ transport and cAMP formation was investigated in immunodissected rabbit kidney connecting and cortical collecting tubule cells grown to confluency on permeable supports. 2. The cyclo-oxygenase inhibitor indomethacin dose-dependently (IC50 = 18 nM) reduced the net apical-to-basolateral Ca2+ transport by 57%. Inhibition was reversed in medium obtained from monolayers incubated in the absence of indomethacin. 3. HPLC analysis following incubation with 14C-labelled arachidonic acid revealed the presence of a wide variety of radiolabelled prostanoids in both the apical and basolateral media. These findings are compatible with the endogenous production and subsequent release of stimulatory prostanoids. 4. The inhibitory action of indomethacin was reversed by the addition of the prostanoids PGE1, PGE2 and PGA2, but not PGD2, PGF2 alpha, the stable PGI2 analogue cicaprost or the thromboxane A2 mimetic U-46619. PGE2 stimulated transepithelial Ca2+ transport dose dependently (EC50 = 3 nM), irrespective of the compartment of which it was added. The stimulatory effect of PGE2 was paralleled by increased cAMP formation, suggesting the apical and basolateral presence of stimulatory prostanoid receptors EP2 and/or EP4. 5. Sulprostone, an analogue selective for EP1 and EP3 receptors, inhibited transepithelial Ca2+ transport in indomethacin-treated monolayers only when applied basolaterally, suggesting the exclusive presence of inhibitory EP receptors on the basolateral membrane. 6. The percentage by which parathyroid hormone and arginine vasopressin increased both transepithelial Ca2+ transport and cAMP formation was dramatically increased in indomethacin-inhibited cells as compared with control cells, demonstrating that indomethacin unmasks the actions of these hormones to their full extent.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Animals; Arachidonic Acid; Arginine Vasopressin; Biological Transport; Calcium; Cells, Cultured; Chromatography, High Pressure Liquid; Cyclic AMP; Cyclooxygenase Inhibitors; Dinoprostone; Eicosanoids; Epoprostenol; Indomethacin; Kidney Tubules; Models, Biological; Parathyroid Hormone; Prostaglandin Endoperoxides, Synthetic; Prostaglandins; Rabbits; Receptors, Prostaglandin; Thromboxane A2; Virulence Factors, Bordetella

[3H]PGE2 and [3H]PGF2 alpha were shown to bind with similar binding capacity and dissociation constants to bovine aorta endothelial cells. The similarity in the binding parameters suggests that both agonists may bind to the same binding site. Displacement of [3H]PGE2 performed with PGE2, PGF2 alpha or U-46619, a thromboxane agonist, shows that all three prostanoids displaced the bound [3H]PGE2 with comparable potency (IC50 = 10(-7) M). These results indicated that the three different prostanoids, which serve as specific agonists to different prostanoid receptors, also compete for the same binding site in bovine endothelial cells with similar affinity. Comparison of the displacement of [3H]PGE2 or [3H]PGF2 alpha by a number of prostaglandin agonists and antagonists further supports the notion that the natural prostanoids bind with similar affinities to the same binding site. Thus, sulprostone, an EP1/EP3 agonist, displaced bound [3H]PGE2 and [3H]PGF2 alpha with IC50 of about 10(-7) M. On the other hand, thromboxane antagonists (BAY u-3405 and GR-32191B), EP1 specific antagonist (SC-19220) EP1/DP antagonist (AH-6809) and iloprost, a stable prostacyclin agonist, failed to displace bound [3H]PGE2 or [3H]PGF2 alpha at a concentration range of 10(-9)-10(-6) M. Gradual increase of sodium fluoride (NaF), a general activator of G binding proteins, or incubation of permeabilized cells with GTP gamma S resulted in a decrease in [3H]PGE2 binding, suggesting that the binding site represents a low-affinity common prostanoid receptor which, similar to other prostanoid receptors, is probably coupled with G binding proteins.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Animals; Aorta; Binding Sites; Biphenyl Compounds; Carbazoles; Cattle; Cells, Cultured; Dibenz(b,f)(1,4)oxazepine-10(11H)-carboxylic acid, 8-chloro-, 2-acetylhydrazide; Dinoprostone; Endothelium, Vascular; Epoprostenol; Heptanoic Acids; Iloprost; Prostaglandin Endoperoxides, Synthetic; Prostaglandins; Sulfonamides; Thromboxane A2; Thromboxanes; Xanthenes; Xanthones

Interleukin-6 (IL-6) is a cytokine involved in the differentiation of B-cells to antibody secreting plasma cells, the activation of T-cells, and the stimulation of hepatocyte production of acute phase proteins. Because of the pro-inflammatory effects of this cytokine, we investigated the ability of the fatty acid arachidonic acid (AA) to regulate the release of IL-6 from rat resident peritoneal macrophages (M phi) in vitro. AA (0.5-16 microM) stimulated IL-6 release during a 4 h incubation period in a biphasic manner, with 4 microM AA generating a peak of IL-6 release (3-5-fold). AA (0.5-16 microM) also induced an increasing release of the AA metabolite thromboxane B2 (TXB2). The AA-induced release of IL-6 occurred within 1-2 h of incubation, whereas TXB2 concentrations were elevated within 5 min of AA treatment. The TX synthetase inhibitor CGS 12970 (4.0 microM and 40.0 microM) effectively blocked the generation of TXB2, but increased prostacyclin (PGI2) generation and potentiated the release of IL-6. In addition, PGI2, as well as the PGI2 agonists iloprost and cicaprost, stimulated IL-6 release from M phi by greater than 5-fold over vehicle-treated basal levels. These data suggest that PGI2 (but not TXA2) is involved in AA-induced IL-6 release from peritoneal M phi.

Topics: Animals; Arachidonic Acid; Epoprostenol; Iloprost; In Vitro Techniques; Interleukin-6; Kinetics; Macrophages, Peritoneal; Male; Prostaglandins, Synthetic; Pyridines; Rats; Thromboxane A2; Thromboxane-A Synthase