15-hydroxy-11-alpha-9-alpha-(epoxymethano)prosta-5-13-dienoic-acid and Bronchial-Hyperreactivity

Asthma is now thought to be a chronic inflammatory disease of the airways. The roles of prostanoids, thromboxane A2 (TXA2) and the prostaglandins (PGs) in the pathogenesis and pathophysiology of asthma have fostered a wealth of studies but remain controversial. TXA2 and the bronchoconstrictor PGs, PGD2 and PGF2 alpha, are generated in greater amounts in asthmatic than in normal subjects. TXA2 is a potent constrictor of airway smooth muscle, an inducer of acetylcholine release and of airway microvascular leakage. It may participate in the thickening and the remodeling of the airway wall which may contribute to the airway hyperresponsiveness, a typical feature of asthma. Strategies for inhibition of TXA2 effects include antagonism of the TXA2 receptor (TP receptor) and inhibition of the thromboxane synthase. TP receptor antagonists could block the effects of all the bronchoconstrictor prostanoids because TXA2 as well as the bronchoconstrictor PGs act through activation of lung TP receptor. The recent development of specific and potent TP receptor antagonists and inhibitors of thromboxane synthase has provided tools to assess the role of TXA2 and broncho-constrictor PGs in the pathophysiology of asthma.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Airway Resistance; Asthma; Bronchial Hyperreactivity; Clinical Trials as Topic; Humans; Lung; Muscle Contraction; Muscle, Smooth; Prostaglandin Endoperoxides, Synthetic; Prostaglandins; Receptors, Thromboxane; Thromboxane A2; Thromboxane-A Synthase; Vasoconstrictor Agents

Chronic inflammation is a hallmark of pulmonary diseases, which leads to lung parenchyma destruction (emphysema) and obstructive bronchiolitis occurring in both chronic obstructive pulmonary disease and asthma. Inflammation is strongly correlated with low intracellular cAMP levels and increase in specific cAMP hydrolyzing activity. The aim of the present study was to investigate the role of the cyclic phosphodiesterase type 4 (PDE4) in human lung and to determine the effects of NCS 613, a new PDE4 inhibitor, on lung inflammation and bronchial hyperresponsiveness. High cAMP-PDE activities were found in the cytosoluble fractions from human lung parenchyma and distal bronchi. PDE4 (rolipram sensitive) represented 40% and 56% of total cAMP-PDE activities in the above-corresponding tissues. Moreover, PDE4A, PDE4B, PDE4C, and PDE4D isoforms were detected in all three subcellular fractions (cytosolic, microsomal, and nuclear) with differential distributions according to specific variants. Pharmacological treatments with NCS 613 significantly decreased PDE4 activity and reduced IκBα degradation in cultured parenchyma, both of which are usually correlated with a lower inflammation status. Moreover, NCS 613 pretreatment potentiated isoproterenol-induced relaxations in human distal bronchi, while reducing TNF-α-induced hyperresponsiveness in cultured bronchi, as assessed in the presence of methacholine, U-46619, or histamine. This reducing effect of NCS 613 on human bronchi hyperresponsiveness triggered by TNF-α was related to a lower expression level of PDE4B and PDE4C, as well as a downregulation of the phosphorylated forms of p38-MAPK, CPI-17, and MYPT-1, which are known to control tone. In conclusion, specific PDE4 inhibitors, such as NCS 613, may represent an alternative and isoform-specific approach toward reducing human lung inflammation and airway overreactivity.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Anti-Inflammatory Agents; Asthma; Bronchi; Bronchial Hyperreactivity; Bronchoconstrictor Agents; Cell Fractionation; Cell Separation; Cyclic AMP; Cyclic Nucleotide Phosphodiesterases, Type 4; Histamine; Humans; Immunohistochemistry; Intracellular Signaling Peptides and Proteins; Isoenzymes; Methacholine Chloride; Middle Aged; Muscle Proteins; Myosin-Light-Chain Phosphatase; p38 Mitogen-Activated Protein Kinases; Phosphodiesterase 4 Inhibitors; Phosphoprotein Phosphatases; Pneumonia; Rolipram; Tissue Culture Techniques; Tumor Necrosis Factor-alpha; Vasoconstrictor Agents

Respiratory pathology research is limited by the number of appropriate multicellular models suitable for studying mechanical properties and signaling pathways that are involved in airway responsiveness. In this study, the electrophysiological and pharmacomechanical properties of organ-cultured explants derived from normal guinea pig bronchi and trachea were investigated. The explants maintained their basic histological phenotype but became hyperreactive to excitatory (muscarinic, histaminergic, serotinergic, and thromboxane receptor agonists, 60 mM KCl) and inhibitory (norepinephrine, isoproterenol) stimuli within the first 3 days in culture, with or without serum in the culture medium. Indomethacin pretreatment did not modify the spasmogen responses of the explant. The onset of this intrinsic overreactivity was highly dependent on the initial presence of epithelium, took 3 days to reach its maximum, and lasted over several days (days 3 to 7). Removal of Ca2+ from the bathing solution initially normalized the inotropic responses of the cultured versus freshly isolated airway tissues. However, the responses to repetitive carbachol challenges in the absence of Ca2+ displayed a slower inactivation in the cultured explants compared to fresh tissues. Smooth muscle resting membrane potential and potassium-induced depolarizations were unaffected by organ culture. Immunohistochemical analyses revealed the presence of apoptotic bodies in the submucosa and epithelial layers, but none in the smooth muscle layer of cultured airways. These functional and histological findings may prove useful in understanding signaling processes involved in tissue hyperresponsiveness related to asthma.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Animals; Apoptosis; Biomechanical Phenomena; Bronchi; Bronchial Hyperreactivity; Calcium; Calcium Signaling; Carbachol; Electrophysiology; Female; Guinea Pigs; Histamine; Indomethacin; Male; Membrane Potentials; Models, Biological; Norepinephrine; Organ Culture Techniques; Time Factors; Trachea

We examined the role of cyclooxygenase in airway hyperresponsiveness and inflammation after ozone exposure in guinea pigs using a non-selective (indomethacin) and a selective (JTE-522) cyclooxygenase-2 inhibitor. Spontaneously breathing guinea pigs were exposed to ozone (3 ppm) for 2 h after treatment with vehicle, indomethacin (10 mg/kg) or JTE-522 (10 mg/kg). Airway responsiveness to inhaled histamine (PC(200)) and bronchoalveolar lavage were assessed before, immediately and 5 h after ozone exposure. Ozone caused a significant airway hyperresponsiveness immediately after exposure, which persisted after 5 h. Neither JTE-522 nor indomethacin affected airway hyperresponsiveness immediately after ozone exposure, but significantly attenuated airway hyperresponsiveness 5 h after exposure, suggesting that cyclooxygenase-2 may participate in the late phase of airway hyperresponsiveness but not in the early phase. Ozone caused a significant increase in the concentration of prostaglandin E(2) and thromboxane B(2) in bronchoalveolar lavage fluid immediately after exposure, which decreased to the basal level 5 h after exposure. This increase in prostaglandin E(2) and thromboxane B(2) was significantly inhibited by JTE-522. An expression of cyclooxygenase-2 was detected not only after ozone exposure but also before, and there was no difference in the number of cyclooxygenase-2-positive cells at any time point. An exogenously applied thromboxane A(2) mimetic, U-46619 (10(-5) M), induced airway hyperresponsiveness 5 h after inhalation, but not immediately or 3 h after inhalation. These data suggest that cyclooxygenase-2 may be constitutively expressed before ozone exposure in guinea pig airway and may synthesize prostaglandin E(2) and thromboxane A(2) transiently under ozone stimulation and that thromboxane A(2) may, in turn, induce the late phase of airway hyperresponsiveness.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Administration, Inhalation; Animals; Benzenesulfonates; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cell Count; Cyclooxygenase 2; Cyclooxygenase 2 Inhibitors; Cyclooxygenase Inhibitors; Dinoprostone; Guinea Pigs; Histamine; Immunohistochemistry; Isoenzymes; Male; Oxazoles; Oxidants, Photochemical; Ozone; Prostaglandin-Endoperoxide Synthases; Prostaglandins; Thromboxane A2

The aim was to determine the time courses for the changes in airway function, airway reactivity, influx of inflammatory cells and levels of the pro-inflammatory cytokines, interleukin (IL)-5 and IL-8 in bronchoalveolar lavage fluid (BALF), and the plasma levels of cortisol and ACTH after antigen challenge to determine whether a temporal link could be established between these events.. Airway function was measured as specific airway conductance (sGsw) in conscious ovalbumin (OvA)-sensitized guinea pigs using whole body plethysmography at intervals after an inhalation challenge with ovalbumin (0.5% for 10 min). Airway responses to the inhaled spasmogen, U46619 (30 ng/ml, 60 s), were measured at 3, 6 and 24 h after challenge. In separate animals, bronchoalveolar lavage fluid (BALF) was obtained after anaesthetic overdose either before challenge or at 1, 3, 6, 12, or 24 h after OvA challenge. Total and differential cell counts of eosinophils and neutrophils were performed on BALF and levels of IL-5 and IL-8 determined by scintillation proximity assays and ELISA, respectively. Plasma cortisol and ACTH levels were determined by RIA kits in blood removed by cardiac puncture at intervals after challenge.. An early phase bronchoconstriction occurred which resolved by 3 h and was followed by a late phase between 17 and 24 h. Airway hyperresponsiveness to inhaled U46619, was evident at 3, 6 and 24 h after antigen challenge. Increased IL-5[BALF] was observed by 60 min post challenge implicating a performed storage site. In contrast, IL-8[BALF] was not raised until 3 h post challenge. There was a significant infiltration of neutrophils and eosinophils by 3 and 6 h, respectively. IL-5[BALF] further increased up to 24 h, during the appearance of the late phase of bronchoconstriction and whilst eosinophilia was maximal. Plasma cortisol levels were increased 1 and 3 hours after antigen challenge, thereafter returning to baseline levels.. The hyperresponsiveness appears to be dissociated from the appearance of eosinophils in lavage fluid. The early appearance of IL-5, however, could be a trigger for the migration of eosinophils and development of hyper-responsiveness. The increased plasma cortisol levels occurring after antigen challenge were presumably due to the stress involved and these would be expected to exert an endogenous anti-inflammatory effect.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Adrenocorticotropic Hormone; Airway Resistance; Animals; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Bronchoconstriction; Cytokines; Guinea Pigs; Hydrocortisone; In Vitro Techniques; Interleukin-5; Interleukin-8; Leukocytes; Lung; Male; Ovalbumin; Plethysmography, Whole Body; Time Factors

The physiological and pharmacological consequences of repeated aero-allergen challenge have not been previously characterized in conscious, sensitized guinea-pigs.. This study was undertaken to compare the effects of two anti-inflammatory compounds, dexamethasone and Ro 20- 1724, on an acute and chronic airway inflammation, in terms of airway function, reactivity and leucocyte infiltration.. Sensitized guinea-pigs received eight saline or ovalbumin (OvA) inhalation exposures over 4 weeks and either vehicle, the type 4 PDE inhibitor, Ro 20-1724 (3 mgkg(-1)), or dexamethasone (1.5 mg/kg(-1)), 30 min before and 6 h after each challenge. Airway function of the conscious animal (sGaw) was monitored over the duration of the first and final OvA challenge. Airway reactivity to the thromboxane mimetic, U46619, was also determined following the final OvA exposure as was the leucocyte infiltration.. The first antigen challenge induced a large early (0-3h) and smaller late (17-24h) bronchoconstrictor response. Neither phase was affected by the drug treatments. The final OvA challenge induced early and late phase bronchoconstrictor responses but of similar magnitude. The late phase was also significantly prolonged. Ro 20-1724 and dexamethasone significantly attenuated both phases. Airway reactivity to the inhaled thromboxane mimetic, U46619, was also significantly enhanced at 120h after the final OvA exposure in contrast to the saline challenged group. This hyperreactivity was attenuated by Ro 20-1724 and dexamethasone. Bronchoalveolar lavage after repeated OvA exposures revealed eosinophilia which was attenuated by Ro 20-1724 and dexamethasone.. This model demonstrates differential airway responses to acute and chronic antigen challenge. Repeated administration of dexamethasone and Ro 20-1724 with each OvA exposure attenuated all of the chronic inflammatory responses: early and late phase responses, hyperreactivity and eosinophilia.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; 3',5'-Cyclic-AMP Phosphodiesterases; 4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone; Administration, Inhalation; Animals; Antigens; Bronchial Hyperreactivity; Cell Count; Cyclic Nucleotide Phosphodiesterases, Type 4; Dexamethasone; Dose-Response Relationship, Drug; Dose-Response Relationship, Immunologic; Eosinophilia; Eosinophils; Glucocorticoids; Guinea Pigs; Inflammation; Macrophages; Male; Neutrophils; Ovalbumin; Phosphodiesterase Inhibitors; Vasoconstrictor Agents

Lipopolysaccharide (LPS) and interleukin (IL)-1beta have been reported to induce airway hyperresponsiveness in several animal models. This study investigated the effect of LPS or IL-1beta on bradykinin-induced human isolated bronchi contraction. LPS (100 ng x mL(-1) for 3-6 h) and IL-1beta (3x10(-10) and 3x10(-9) M for 20 min to 3 h) time-dependently potentiated bradykinin-induced contraction. This contraction was abolished, as in control experiments, by indomethacin (10(-6) M) or by the thromboxane (Tx) receptor antagonist GR 32191 but not by the cyclo-oxygenase-2 inhibitor, CGP28238. In contrast, the Tx mimetic U46619-induced contraction of human bronchi was not enhanced IL-1beta pretreatment. In the presence of GR 32191 (10(-6) M), bradykinin induced a prostanoid dependent relaxation that was not significantly modified by IL-1beta pretreatment. Determination of prostanoids in the organ bath fluid showed that bradykinin induced TxB2, the stable metabolite of TxA2, and 6-keto prostaglandin F1alpha, the stable metabolite of PGI2, release. Only TxA2 release was potentiated by IL-1beta. Taken together our results suggest that interleukin-1beta (1-3 h)-induced potentiation of the effect of bradykinin is linked to an increased activity of thromboxane synthase and, in turn, to increased thromboxane synthesis.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; 6-Ketoprostaglandin F1 alpha; Aged; Biphenyl Compounds; Bradykinin; Bronchi; Bronchial Hyperreactivity; Bronchoconstriction; Cyclooxygenase Inhibitors; Dose-Response Relationship, Drug; Heptanoic Acids; Humans; In Vitro Techniques; Indomethacin; Interleukin-1; Lipopolysaccharides; Middle Aged; Receptors, Thromboxane; Thromboxanes

1. The aim of this study was to determine the effects of inhibitors of phosphodiesterase (PDE) on the early and late phase bronchoconstriction in sensitized, conscious guinea-pigs and the subsequent development of acute airway hyperreactivity to the inhaled thromboxane mimetic, U46619, and leukocyte infiltration following ovalbumin (OvA) challenge. 2. Following an inhalation challenge with OvA, there was an early bronchoconstriction which peaked at 15 min with recovery after 3-4 h. A late phase bronchoconstriction occurred between 17 and 24 h after challenge. The PDE 4 inhibitors, Ro 20-1724 (3 mg kg-1, i.p.) and rolipram (1 mg kg-1, i.p.) administered 30 min before and 6 h after antigen challenge (double dosing regimen), did not affect the development of the early or late phase responses. 3. Seventeen to twenty four hours following an acute OvA or saline challenge, a consistently greater bronchoconstrictor response to inhaled U46619 was observed in the OvA challenged group. This increase in responsiveness was significantly attenuated by the administration of Ro 20-1724 and rolipram 30 min before and 6 h after antigen challenge (P < 0.05); this was not attributable to a residual bronchodilator effect of these compounds. There was a trend towards inhibition of the hyperreactivity to U46619 by aminophylline but not by the PDE3 inhibitors, siguazodan or SKF 95654. 4. Aminophylline, rolipram and Ro 20-1724 when administered as the double dose regimen attenuated the rise in macrophages, eosinophils and neutrophils recovered in bronchial lavage fluid 17 to 24 h after antigen challenge. 5. The dose of Ro 20-1724 given at 6 h post challenge was essential for attenuation of airway hyperreactivity and to protect against leukocyte influx. 6. In summary, aminophylline, rolipram and Ro 20-1724 have anti-inflammatory effects against antigen-induced airway leukocyte infiltration. Rolipram and Ro 20-1724 additionally attenuated the development of acute airway hyperreactivity, effects which are probably mediated through inhibition of PDE type 4. A dose of PDE inhibitor 6 h after the antigen challenge appears to be essential to achieve this protection. Inhibitors of PDE type 3 were generally without effect. However, there was no effect of rolipram or Ro 20-1724 on the development of either the early or late phase type responses.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; 4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone; Aminophylline; Animals; Bronchial Hyperreactivity; Guinea Pigs; Leukocytes; Male; Ovalbumin; Phosphodiesterase Inhibitors; Prostaglandin Endoperoxides, Synthetic; Pyrrolidinones; Rolipram; Thromboxane A2

Antigen-stimulated contraction and release of chemical mediators were examined in saline- or Sephadex-treated rat lung parenchymal strips. Sephadex treatment caused eosinophilia in the blood and the lung tissue. Antigen challenge of the isolated parenchymal strips in Sephadex-treated rat was followed by passive sensitization, resulted in an augmented contraction and elevated releases of thromboxane (TX) B2 and peptide-leukotrienes (p-LTs) in bath fluid compared with those of saline-treated control. Although 5-hydroxytryptamine (5-HT) and histamine were significantly released after antigen challenge, the levels were not different between saline- and Sephadex-treated groups. DP-1904, a selective thromboxane synthetase inhibitor, and methysergide but not atropine significantly reduced the augmented contraction and inhibited the elevated TXB2 release in the Sephadex-treated group. Similar increased contraction and the elevated TXB2 release above were observed when Sephadex-treated rat lung strips were stimulated by exogenous 5-HT and LTD4. These augmented contractions were closely correlated with the increase in TXB2 level (r = 0.83; p < 0.01). In addition, contraction to U-46619, a thromboxane mimetic, was significantly greater in Sephadex-treated rat lung strips. Our results indicate that the ability of Sephadex-treated rat lung tissue to synthesize newly generated mediators such as TXA2 and p-LTs is increased, and the spasmogenic susceptibility of the lung tissue to TXA2 itself is modified by Sephadex treatment, suggesting these are due to the augmented contraction in an established hyperresponsiveness state induced by Sephadex.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Animals; Atropine; Bronchial Hyperreactivity; Dextrans; Dose-Response Relationship, Drug; Imidazoles; Leukotriene D4; Lung; Male; Methysergide; Prostaglandin Endoperoxides, Synthetic; Pulmonary Eosinophilia; Rats; Rats, Sprague-Dawley; Tetrahydronaphthalenes; Thromboxane A2; Vasoconstrictor Agents

The effect of the anti-allergic drug azelastine, 4-(p-chlorobenzyl)-2-(hexahydro-1-methyl-1H-azepine-4-yl)-1-(2H)-phth alazione), on airway hyperresponsiveness induced by immunologically stimulated pulmonary alveolar macrophages was investigated in canine bronchial segments under isometric conditions in vitro. Macrophages stimulated with anti-dinitrophenyl immunoglobulin E (IgE) antibody and dinitrophenyl-human serum albumin potentiated the contractile responses to electrical field stimulation at all frequencies, an effect that was abolished by azelastine (3 x 10(-5) M). In contrast, azelastine had no effect on the potentiation of the contractile responses to electrical stimulation by U46619, a thromboxane A2 mimetic. The IgE-mediated release of thromboxane A2 from macrophages was inhibited by azelastine in a concentration-dependent fashion, the maximal decrease and the concentration required to produce a half-maximal effect being 84 +/- 6% (P < 0.001) and 16 microM, respectively. These results suggest that azelastine may attenuate macrophage-induced parasympathetic airway hyperresponsiveness through an inhibition of the release of thromboxane A2.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Animals; Bronchial Hyperreactivity; Dogs; Electric Stimulation; Female; Histamine H1 Antagonists; Immunoglobulin E; In Vitro Techniques; Isometric Contraction; Macrophage Activation; Macrophages, Alveolar; Male; Parasympathetic Nervous System; Phthalazines; Prostaglandin Endoperoxides, Synthetic; Serum Albumin; Synaptic Transmission; Thromboxane A2; Vasoconstrictor Agents

The effect of a novel thromboxane A2 receptor antagonist, BAY-u-3405, on experimental allergic airway and skin reactions was studied in vivo. At doses of 3-30 mg/kg BAY-u-3405 clearly inhibited the U-46619-induced increase in respiratory resistance (Rrs) in guinea pigs. BAY-u-3405 at doses of 3 and 30 mg/kg inhibited the aerosolized antigen-induced biphasic increase in respiratory resistance in guinea pigs. Moreover, BAY-u-3405 inhibited repeated aeroantigen-induced airway hyperactivity and airway inflammation in mice. In IgE antibody-mediated biphasic skin reactions in mice, both immediate and late-phase reactions were inhibited by 10 mg/kg of BAY-u-3405. These results demonstrate the efficacy of BAY-u-3405 on the antigen-induced late-phase reactions in the airway and skin in guinea pigs and mice, and antigen-induced airway hyperactivity in mice.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Animals; Bronchial Hyperreactivity; Carbazoles; Dermatitis; Guinea Pigs; Hypersensitivity; Immunoglobulin E; Male; Mice; Mice, Inbred BALB C; Platelet Aggregation Inhibitors; Prostaglandin Endoperoxides, Synthetic; Receptors, Thromboxane; Sulfonamides; Thromboxane A2

The anti-asthmatic effects of CS-518 (sodium 2-(1-imidazolylmethyl)-4,5-dihydrobenzo[b]thiophene-6-carboxylate) , a specific thromboxane A2 (TXA2) synthase inhibitor, were investigated in the ovalbumin-sensitized guinea pig asthmatic model. Although CS-518 slightly inhibited (about 25%) whole bronchoconstriction, it significantly inhibited the antigen-induced bronchoconstriction mediated by slow-reacting substance of anaphylaxis (SRS-A), which was not reduced by chlorpheniramine, a histamine H1 antagonist. On the other hand, indomethacin, a cyclooxygenase inhibitor, potentiated the SRS-A-mediated constriction. CS-518 strongly, and indomethacin slightly, suppressed the leukotriene D4-induced bronchoconstriction. CS-518 clearly inhibited the antigen-induced airway hyperresponsiveness, but this compound had no effect on the airway hyperresponsiveness induced by U-46619, a TXA2-mimetic agent, and propranolol. These results suggest that CS-518 suppresses the development of bronchoconstriction and airway hyperresponsiveness in asthmatic models by inhibition of TXA2 synthesis with the concomitant increase in bronchodilating prostaglandins such as prostaglandin E2 and prostaglandin I2.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Animals; Asthma; Bronchial Hyperreactivity; Bronchoconstriction; Chlorpheniramine; Disease Models, Animal; Guinea Pigs; Indomethacin; Male; Methacrylates; Ovalbumin; Propranolol; Prostaglandin Endoperoxides, Synthetic; SRS-A; Thiophenes; Thromboxane A2; Thromboxane-A Synthase; Vasoconstrictor Agents