dinoprost and carboprostacyclin

Cyclooxygenase (COX)-2 is among the endothelial genes upregulated by uniform laminar shear stress (LSS), characteristically associated with atherosclerotic lesion-protected areas. We have addressed whether the induction of COX-2-dependent prostanoids in endothelial cells by LSS plays a role in restraining endothelial tumor necrosis factor (TNF)-alpha generation, a proatherogenic cytokine, through the induction of heme oxygenase-1 (HO)-1, an antioxidant enzyme. In human umbilical vein endothelial cells (HUVECs) exposed to steady LSS of 10 dyn/cm(2) for 6 hours, COX-2 protein was significantly induced, whereas COX-1 and the downstream synthases were not significantly modulated. This was associated with significant (P<0.05) increase of 6-keto-prostaglandin (PG)F(1alpha) (the hydrolysis product of prostacyclin), PGE(2), and PGD(2). In contrast, TNF-alpha released in the medium in 6 hours (3633+/-882 pg) or detected in cells lysates (1091+/-270 pg) was significantly (P<0.05) reduced versus static condition (9100+/-2158 and 2208+/-300 pg, respectively). Coincident induction of HO-1 was detected. The finding that LSS-dependent reduction of TNF-alpha generation and HO-1 induction were abrogated by the selective inhibitor of COX-2 NS-398, the nonselective COX inhibitor aspirin, or the specific prostacyclin receptor (IP) antagonist RO3244794 illuminates the central role played by LSS-induced COX-2-dependent prostacyclin in restraining endothelial inflammation. Carbacyclin, an agonist of IP, induced HO-1. Similarly to inhibition of prostacyclin biosynthesis or activity, the novel imidazole-based HO-1 inhibitor QC15 reversed TNF-alpha reduction by LSS. These findings suggest that inhibition of COX-2-dependent prostacyclin might contribute to acceleration of atherogenesis in patients taking traditional nonsteroidal antiinflammatory drugs (NSAIDs) and NSAIDs selective for COX-2 through downregulation of HO-1, which halts TNF-alpha generation in human endothelial cells.

Topics: 6-Ketoprostaglandin F1 alpha; Aspirin; Atherosclerosis; Benzofurans; Cells, Cultured; Cyclooxygenase 1; Cyclooxygenase 2; Cyclooxygenase Inhibitors; Dinoprost; Dinoprostone; Down-Regulation; Endothelial Cells; Epoprostenol; Heme Oxygenase-1; Humans; Inflammation; Nitrobenzenes; Perfusion; Propionates; Prostaglandin D2; Receptors, Epoprostenol; Receptors, Prostaglandin; Stress, Mechanical; Sulfonamides; Tumor Necrosis Factor-alpha; Up-Regulation

The pharmacological properties of [(3)H]-prostaglandin E(2) ([(3)H]-PGE(2)) binding to washed homogenates of hamster uterus were determined. Scatchard analysis of competition data yielded dissociation constants (K(d)s) of 30.9 +/- 5.6 nM (n = 3) and apparent receptor density (B(max)) of 25.25 +/- 1.89 pmol g(-1) wet weight tissue (74 +/- 8% specific binding). Competition studies yielded the following affinity parameters (K(i)) for various prostanoids: GR63799X = 13 4 nM; PGE(2) = 17 +/- 3 nM; sulprostone = 64 +/- 5 nM; enprostil = 67 +/- 3 nM; misoprostol = 124 +/- 15 nM; cloprostenol = 187 +/- 33 nM; carba-prostacyclin = 260 +/- 167 nM; iloprost = 555 +/- 162 nM; PGF(2 alpha) = 767 +/- 73 nM; PGD(2) > 3560 nM; fluprostenol = 11 790 +/- 2776 nM; RS93520 = 21 558 +/- 14 228 nM. These data closely matched the pharmacological profile of previously described EP(3) receptors such as in bovine corpus luteum (BCLM) and the cloned mammalian EP(3) receptors. The high correlation between the current hamster uterus pharmacology data vs the EP(3) receptor binding in BCLM (r = 0.94; P < 0.0001), vs cloned human EP(3) receptor (r = 0.94, P < 0.0001), vs the cloned mouse EP(3) receptor binding (r = 0.78; P < 0.002), vs cloned rat EP(3) receptor (r = 0.9, P < 0.0004), and vs EP(3) receptor-mediated functional responses (r = 0.72, P < 0.02) substantiated the conclusion that the hamster uterus contains EP(3) receptor binding sites.

Topics: Animals; Binding Sites; Biphenyl Compounds; Bridged Bicyclo Compounds, Heterocyclic; Cattle; Cloprostenol; Cricetinae; Dinoprost; Dinoprostone; Enprostil; Epoprostenol; Fatty Acids, Unsaturated; Female; Hydantoins; Hydrazines; Iloprost; Latanoprost; Misoprostol; Prostaglandins; Prostaglandins E, Synthetic; Prostaglandins F, Synthetic; Receptors, Prostaglandin E; Receptors, Prostaglandin E, EP3 Subtype; Tritium; Uterus

To determine whether prostanoids, nonsteroidal anti-inflammatory drugs (NSAIDs), and members of the tumor necrosis factor superfamily can regulate placental secretion of interleukin-6 (IL-6) and whether labor influences any such effects.. Villous fragments of term, human placenta were kept in culture for up to 4 hours, and IL-6 concentrations were measured in the supernatant. We assessed the effects of the following prostanoids: PGE(2), PGF(2alpha), thromboxane A(2) mimetic (U-46619), and carbacyclin, a stable prostacyclin analogue (all at 1 microM); NSAIDs: indomethacin (150 microM) or nimesulide (100 microM); and Fas ligand (5 ng/mL).. Secretion (mean +/- standard error) of IL-6 was, for control conditions, 1.92 +/- 0.28 fmol/mg wet weight per 3 hours; for PGE(2), 3.57 +/- 0.29 fmol/mg wet weight per 3 hours, P <.01; and for carbacyclin, 3.11 +/- 0.44 fmol/mg wet weight per 3 hours, P <.01. Incubation with PGF(2alpha) or the thromboxane A(2) analogue, U46619, had no effect on IL-6 secretion under these conditions. Fas ligand stimulated IL-6 secretion (3.06 +/- 0.38 fmol/mg wet weight per 3 hours, P <.05). Labor did not alter the effects of prostanoids or FasL. The effects of NSAIDs were assessed over 4 hours. Secretion (median, interquartile range) was, under control conditions 3.26, 2.83-6.23 fmol/mg wet weight per 4 hours, with indomethacin 1.4, 1.28-3.21 (P <.05), and with nimesulide 0.75, 0.50-1.56 fmol/mg wet weight per 4 hours. The magnitude of the effect of Fas ligand in the presence of NSAIDs depended on whether the placentas were delivered before or after labor.. Prostanoids, NSAIDs, and the Fas ligand regulate placental IL-6 secretion. Although the effects of individual agents did not vary with the presence or absence of labor, modulation of IL-6 secretion by labor became apparent when agents were combined.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Anti-Inflammatory Agents, Non-Steroidal; Dinoprost; Dinoprostone; Drug Interactions; Epoprostenol; Fas Ligand Protein; Female; Humans; Interleukin-6; Labor, Obstetric; Membrane Glycoproteins; Models, Biological; Placenta; Pregnancy; Prostaglandins; Thromboxane A2; Tumor Necrosis Factor-alpha

Prostanoids are arachidonic acid (AA) metabolites derived from the cyclooxygenase (COX1 and COX2 isozymes) pathway and are involved in signal transduction pathways activated by distinct ILs. Although COX1 is the constitutive isoform of COX, IL-1beta is a potent inducer of COX2 expression in distinct cell types. This study was designed to determine whether cyclooxygenases could mediate endogenous cytokine regulation in rat progenitor Leydig cells. COX and IL (IL-1alpha, IL-1beta, and IL-6) mRNAs were measured by PCR and real-time PCR analyses, respectively. COX function was assessed using COX activity inhibitors: indomethacin (INDO; COX1 and COX2 inhibitor) and NS-398 (COX2 selective inhibitor). Our data indicate that endogenous progenitor COX1 mRNA levels are low and are not regulated by IL-1beta. In contrast, COX2 mRNA is induced by IL-1beta at 6, 9, and 24 h. IL-1beta induction of IL mRNAs was in part significantly impaired in the presence of INDO or NS-398. Among the prostanoids tested, prostaglandin E(2) (PGE(2)), PGF(2alpha), and carbaprostacyclin reversed the INDO inhibition of IL production. PGs alone have no (IL-1alpha and IL-1beta) or a modest (IL-6) effect on IL mRNA levels. PGE(2), PGF(2alpha), and PGI(2) measurements show that IL-1beta treatment significantly increases progenitor Leydig cell production of these PGs. Taken together, our data demonstrate that this COX2 cascade is a regulator of cytokines in Leydig progenitors.

Topics: Animals; Cyclooxygenase 1; Cyclooxygenase 2; Cyclooxygenase 2 Inhibitors; Cyclooxygenase Inhibitors; Dinoprost; Dinoprostone; Epoprostenol; Gene Expression Regulation; Indomethacin; Interleukin-1; Interleukin-6; Isoenzymes; Leydig Cells; Male; Membrane Proteins; Polymerase Chain Reaction; Prostaglandin-Endoperoxide Synthases; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Stem Cells

The uterus is a target for prostaglandins, especially at the end of gestation. Whether potassium channels are involved in the effect of prostaglandins is not clear. The aim of this study was to find out.. Concentration-response relationships to prostaglandins (prostaglandin F2alpha, prostaglandin E2, and prostaglandin I2 [carbacyclin]; 10(-10) mol/L-10(-4) mol/L) were studied in isolated uterine rings from mid pregnancy (day 14) and late pregnancy (day 21) rats (Krebs solution, 5% CO2 in air, 37 degrees C; pH, 7.4). Rings were incubated for 30 minutes with either solvent or adenosine triphosphate-sensitive potassium channel inhibitor or opener glibenclamide and levcromakalim or with calcium-sensitive potassium channel inhibitor or opener NS1619 and iberiotoxin, respectively. The changes in integral activity were compared after each concentration of the agent and were expressed as a percent of the basal integral activity.. The increases in spontaneous contractile activity induced by prostaglandin E2 and carbacyclin, but not prostaglandin F2alpha, were statistically significantly higher in tissues from late pregnancy versus mid pregnancy rats and were not affected by any of the K-channel openers or inhibitors.. Adenosine triphosphate-sensitive and calcium-sensitive potassium channels are not involved in the effect of prostaglandin F2alpha, prostaglandin E2, and prostaglandin I2 on pregnant rat uterus.

Topics: Animals; Dinoprost; Dinoprostone; Dose-Response Relationship, Drug; Epoprostenol; Female; Gestational Age; In Vitro Techniques; Muscle, Smooth; Osmolar Concentration; Potassium Channels; Pregnancy; Pregnancy, Animal; Rats; Rats, Sprague-Dawley; Uterine Contraction; Uterus

Prostaglandin endoperoxide H synthases and their arachidonate products have been implicated in modulating angiogenesis during tumor growth and chronic inflammation. Here we report the involvement of thromboxane A(2), a downstream metabolite of prostaglandin H synthase, in angiogenesis. A TXA(2) mimetic, U46619, stimulated endothelial cell migration. Angiogenic basic fibroblast growth factor (bFGF) or vascular endothelial growth factor (VEGF) increased TXA(2) synthesis in endothelial cells three- to fivefold. Inhibition of TXA(2) synthesis with furegrelate or CI reduced HUVEC migration stimulated by VEGF or bFGF. A TXA(2) receptor antagonist, SQ29,548, inhibited VEGF- or bFGF-stimulated endothelial cell migration. In vivo, CI inhibited bFGF-induced angiogenesis. Finally, development of lung metastasis in C57Bl/6J mice intravenously injected with Lewis lung carcinoma or B16a cells was significantly inhibited by thromboxane synthase inhibitors, CI or furegrelate sodium. Our data demonstrate the involvement of TXA(2) in angiogenesis and development of tumor metastasis.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Animals; Benzofurans; Bridged Bicyclo Compounds, Heterocyclic; Chemotaxis; Dinoprost; Dinoprostone; Endothelial Growth Factors; Endothelium, Vascular; Enzyme Inhibitors; Epoprostenol; Fatty Acids, Unsaturated; Fibroblast Growth Factor 2; Humans; Hydrazines; Lung Neoplasms; Lymphokines; Male; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Neoplasm Metastasis; Neovascularization, Pathologic; Rats; Receptors, Thromboxane; Thromboxane A2; Thromboxane-A Synthase; Umbilical Veins; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

This study was conducted to determine if high perinatal prostaglandin (PG) and thromboxane (TxA2) levels modified their choroidal vasomotor effects and receptor levels. Both nonperfused (eyecup preparations) and perfused choroidal vessels from saline- or ibuprofen-treated 1-day-old pigs and tissues from adult pigs were used; all prostanoids produced similar vasomotor effects on both preparations. Choroidal PGF2alpha, TxA2, PGI2, and PGD2 levels were higher in the newborn than in adult pigs; injections of ibuprofen (40 mg/kg every 4 h for 48 h) into newborn pigs significantly decreased choroidal levels of all these prostanoids. PGF2alpha and the TxA2 mimetic U-46619 caused less choroidal vasoconstriction and production of inositol 1,4,5-trisphosphate (IP3) in the newborn than in adult pigs. Ibuprofen treatment increased choroidal PGF2alpha vasoconstrictor effects, IP3 production, and receptors, but did not modify response to U-46619. Carbaprostacyclin (PGI2 analog) caused a greater choroidal vasodilatation and adenosine adenosine 3',5'-cyclic monophosphate (cAMP) production in the newborn than in adult pigs; these effects were not modified by ibuprofen. PGD2 did not increase cAMP but caused greater dilatation and nitrite [oxidation product of nitric oxide (NO)] production in the choroid of newborn than of adult pigs, which were decreased to adult levels by ibuprofen and the NO synthase inhibitor N(omega)-nitro-L-arginine. These data suggest that high perinatal PG levels downregulate PGF2alpha receptors and vascular effects but do not modify choroidal responses to TxA2 and PGI2; NO seems to contribute to the vasodilator effects of PGD2.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Aging; Animals; Animals, Newborn; Choroid; Cyclic AMP; Cyclic GMP; Dinoprost; Down-Regulation; Epoprostenol; Ibuprofen; Inositol 1,4,5-Trisphosphate; Nitric Oxide; Nitroarginine; Prostaglandin D2; Prostaglandin Endoperoxides, Synthetic; Prostaglandins; Prostaglandins, Synthetic; Receptors, Prostaglandin; Swine; Thromboxane A2; Vasoconstrictor Agents; Vasodilation

The cellular mechanisms giving rise to the enhanced excitability induced by prostaglandin E2 (PGE2) and carba prostacyclin (CPGI2) in embryonic rat sensory neurons were investigated using the whole cell patch-clamp recording technique. Exposing sensory neurons to 1 microM PGE2 produced a twofold increase in the number of action potentials elicited by a ramp of depolarizing current, but this eicosanoid had no effect on the resting membrane potential or the amplitude of the slow afterhyperpolarization. Characterization of the outward potassium currents in the embryonic sensory neurons indicated that the composition of the total current was variable among these neurons. A steady-state inactivation protocol was used to determine the extent of residual noninactivating current. A conditioning prepulse to +20 mV demonstrated that some of these neurons exhibited only a sustained potassium current with little steady-state inactivation whereas other exhibited some combination of a sustained as well as a rapidly inactivating IA-type current. Treatment with 1 microM PGE2 or 1 microM CPGI2, but not 1 microM prostaglandin F2 alpha (PGF2 alpha) produced a time-dependent suppression of the total potassium current. After a 20-min exposure, PGE2 and CPGI2 inhibited the maximal current obtained at +60 mV by 48 and 40%, respectively. The prostaglandin-induced suppression of the potassium current was not associated with a shift in the voltage dependence for activation. Subtraction of the currents remaining after PGE2 or CPGI2 treatment from their respective control recordings revealed that the prostaglandin-sensitive current had characteristics that were consistent with a sustained-type of potassium current. This idea is supported by the following observation. The steady-state inactivation protocol revealed that for prepulse voltages activating both rapidly inactivating and sustained currents, the relaxation of the current was accelerated after treatment with PGE2 or CPGI2 suggesting the removal of a slower component. This effect was not observed in neurons exhibiting only the sustained type current. These results suggest that pro-inflammatory prostaglandins enhance the excitability of rat sensory neurons, in part, through the suppression of an outward potassium current that may modulate the firing threshold for generation of the action potential.

Topics: Animals; Cells, Cultured; Dinoprost; Dinoprostone; Electrophysiology; Epoprostenol; Membrane Potentials; Neurons, Afferent; Patch-Clamp Techniques; Potassium Channels; Prostaglandins; Rats

1. Comparison of the rank order of potency of the natural prostanoids prostaglandin E2 (PGE2), PGD2, PGF2 alpha and carbaprostacyclin in stimulating cyclic AMP in Jurkat cells is consistent with the presence of an EP receptor. 2. Lack of responsiveness to the EP1/EP3 selective agonist, sulprostone, and the EP2 agonists, butaprost and AH 13205, indicates that this receptor is not of the EP1, EP2 or EP3 subtypes. 3. Inhibition of PGE2-stimulated cyclic AMP by the EP4 antagonist, AH 23848 is non-competitive, unlike the competitive antagonism reported in the pig saphenous vein EP4 preparation. Furthermore, 16,16-dimethyl PGE2 is 100 fold less potent than PGE2 in Jurkat cells, while these agonists are equipotent in the rabbit jugular vein purported EP4 preparation. In addition, 1-OH PGE1, which also is active in the rabbit jugular vein preparation, is inactive in Jurkat cells at concentrations up to 1 x 10(-4) M. These data are not wholly consistent with any adenylate cyclase coupled EP receptor described to date. 4. It is postulated that an EP receptor, positively coupled to adenylate cyclase, with a unique pharmacological profile is present in Jurkat cells.

Topics: Adenylyl Cyclases; Animals; Biphenyl Compounds; Bridged Bicyclo Compounds, Heterocyclic; Cyclic AMP; Dinoprost; Dinoprostone; Epoprostenol; Fatty Acids, Unsaturated; Humans; Hydrazines; Leukemia, T-Cell; Prostaglandin D2; Rabbits; Receptors, Prostaglandin E; Swine; T-Lymphocytes; Tumor Cells, Cultured

Prostacyclin (PGI2) is a potent prostanoid producing various symptoms of inflammation, including an increased sensitivity to noxious stimulation. One component of these PGI2-mediated actions may involve activation or sensitization of sensory neurons to enhance release of neuroactive peptides. We, therefore, examined whether PGI2 and carba prostacyclin (CPGI2), a stable analog of PGI2, could alter the resting and evoked release of the neuropeptides, substance P (SP) and calcitonin gene-related peptide (CGRP) from embryonic rat sensory neurons grown in culture. Treating isolated sensory neurons with CPGI2 (10-1000 nM) for 30 min caused a 3-fold increase in the resting release of both peptides. One nM CPGI2, a concentration that did not alter the resting release, significantly enhanced neuropeptide release evoked by capsaicin, 100 nM bradykinin, or 40 mM KCl. Similarly, 10 nM PGI2 did not alter resting release, but augmented capsaicin-stimulated release of SP and CGRP 2-3 fold. In contrast, prostaglandin F2 alpha was ineffective in altering either resting or capsaicin-evoked peptide release. Our results demonstrate that low concentrations of PGI2 sensitize sensory neurons to other stimuli, whereas higher concentrations evoke release directly. This PGI2-induced augmentation of neuropeptide release may be one mechanism contributing to neurogenic inflammation.

Topics: Animals; Bradykinin; Calcitonin Gene-Related Peptide; Cells, Cultured; Dinoprost; Drug Synergism; Epoprostenol; Evoked Potentials; Female; Neurons, Afferent; Potassium; Rats; Rats, Sprague-Dawley; Stimulation, Chemical; Substance P

FTIR spectra measurements and full optimization curve analysis of their spectra were done to obtain parameters of the OH and C = O stretching vibration bands for intramolecular hydrogen bondings in thromboxane (TX)A2 receptor partial agonist (CTA2), prostaglandin (PG)E2, PGD2, PGF2 alpha, prostacyclin (PGI2) receptor agonist (carbacyclin), and their related compounds in dilute CCl4 solutions. For CTA2, PGE2, PGD2, and PGF2 alpha, cyclic intramolecular hydrogen bonds involving a 15-membered ring similar to that observed for the TXA2 receptor agonist (U-46619) were found between a carboxyl group of the alpha-side chain and a 15-hydroxyl group of the omega-side chain. The arrangement of these side chains was P-shaped, and the percentage of the intramolecular hydrogen-bonded molecules with the 15-membered ring in CCl4 solution showed a high value of ca. 80% for these compounds. In addition, it was found that the cyclic intramolecular hydrogen bonds involving the 13-, 12-, and 12-membered rings in PGE2, PGD2, and PGF2 alpha, respectively, are formed between the carboxyl group of the alpha-side chain and the 11-, 9-, and 9-hydroxyl groups of a cyclopentane ring, respectively, although the percentages of the intramolecular hydrogen-bonded molecules with these membered rings are very small. It was also found that the hydrogen bond is more easily formed in the order of the 11-, 9-, and 15-hydroxyl groups. For carbacyclin, the cyclic intramolecular hydrogen bond involving the 13-membered ring was found between the carboxyl group of the alpha-side chain and the 11-hydroxyl group. The percentage of the intramolecular hydrogen-bonded molecules showed the value of 58% for carbacyclin. On the basis of information on the side-chain conformations in CCl4, we examined the structure-activity relationships for U-46619 in place of TXA2, PGE2, PGD2, PGF2 alpha, and carbacyclin in place of PGI2.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Animals; Bridged Bicyclo Compounds; Carbon Tetrachloride; Dinoprost; Dinoprostone; Dogs; Epoprostenol; Fatty Acids, Monounsaturated; Hydrogen Bonding; Macromolecular Substances; Models, Molecular; Molecular Structure; Prostaglandin D2; Prostaglandin Endoperoxides, Synthetic; Receptors, Thromboxane; Solutions; Spectroscopy, Fourier Transform Infrared; Structure-Activity Relationship; Thromboxane A2

In addition to cAMP production, a transient elevation of intracellular free Ca2+ has been shown to take place in preadipose cells upon stimulation by carbaprostacyclin (cPGI2), both messengers acting in synergy to initiate adipose cell differentiation (Vassaux, G., Gaillard, D., Ailhaud, G., and Négrel, R. (1992) J. Biol. Chem.267, 11092-11097). Further studies reported herein show that this Ca2+ transient is i) elicited by the natural prostaglandin PGI2, ii) independent of the presence of extracellular Ca2+, suggesting a mobilization of Ca2+ from intracellular pools and ii) unaffected by cAMP elevating agents. Moreover, and in contrast to the InsP3-dependent Ca2+ signal evoked by PGF2 alpha, that induced by PGI2 is fully abolished by pretreatment with phorbol esters (EC50: 1-5 nM). Furthermore, experiments designed to empty the Ca2+ pools, using PGI2 or PGF2 alpha as Ca2+ mobilizing agents as well as pretreatments with drugs, allow to conclude that PGI2 mobilizes Ca2+ from an InsP3 sensitive, ryanodine insensitive intracellular pool. Altogether, these results strongly suggest that PGI2 mobilizes Ca2+ from an intracellular store common to that affected by InsP3, by means of a mechanism which remains to be elucidated.

Topics: 1-Methyl-3-isobutylxanthine; Adipose Tissue; Animals; Calcium; Cell Line; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Dinoprost; Enzyme Activation; Epoprostenol; Isoproterenol; Male; Mice; Mice, Inbred C57BL; Mice, Obese; Phorbol Esters; Second Messenger Systems; Stem Cells; Tetradecanoylphorbol Acetate