ABSTRACT
BACKGROUND: The use of inhaled glucocorticoids for persistent asthma causes a temporary reduction in growth velocity in prepubertal children. The resulting decrease in attained height 1 to 4 years after the initiation of inhaled glucocorticoids is thought not to decrease attained adult height. METHODS: We measured adult height in 943 of 1041 participants (90.6%) in the Childhood Asthma Management Program; adult height was determined at a mean (±SD) age of 24.9±2.7 years. Starting at the age of 5 to 13 years, the participants had been randomly assigned to receive 400 µg of budesonide, 16 mg of nedocromil, or placebo daily for 4 to 6 years. We calculated differences in adult height for each active treatment group, as compared with placebo, using multiple linear regression with adjustment for demographic characteristics, asthma features, and height at trial entry. RESULTS: Mean adult height was 1.2 cm lower (95% confidence interval [CI], -1.9 to -0.5) in the budesonide group than in the placebo group (P=0.001) and was 0.2 cm lower (95% CI, -0.9 to 0.5) in the nedocromil group than in the placebo group (P=0.61). A larger daily dose of inhaled glucocorticoid in the first 2 years was associated with a lower adult height (-0.1 cm for each microgram per kilogram of body weight) (P=0.007). The reduction in adult height in the budesonide group as compared with the placebo group was similar to that seen after 2 years of treatment (-1.3 cm; 95% CI, -1.7 to -0.9). During the first 2 years, decreased growth velocity in the budesonide group occurred primarily in prepubertal participants. CONCLUSIONS: The initial decrease in attained height associated with the use of inhaled glucocorticoids in prepubertal children persisted as a reduction in adult height, although the decrease was not progressive or cumulative. (Funded by the National Heart, Lung, and Blood Institute and the National Center for Research Resources; CAMP ClinicalTrials.gov number, NCT00000575.).
Subject(s)
Asthma/drug therapy , Body Height/drug effects , Budesonide/pharmacology , Glucocorticoids/pharmacology , Growth/drug effects , Nedocromil/pharmacology , Administration, Inhalation , Adolescent , Adult , Anti-Asthmatic Agents/pharmacology , Anti-Asthmatic Agents/therapeutic use , Asthma/physiopathology , Budesonide/therapeutic use , Child , Child, Preschool , Female , Follow-Up Studies , Glucocorticoids/therapeutic use , Humans , Intention to Treat Analysis , Male , Nedocromil/therapeutic useABSTRACT
Previously, we have reported sex differences in the cardiac remodeling response to ventricular volume overload whereby male and ovariectomized (OVX) female rats develop eccentric hypertrophy, and intact (Int) female rats develop concentric hypertrophy. In males, this adverse remodeling has been attributed to an initial cascade of events involving myocardial mast cell and matrix metalloproteinase activation and extracellular collagen matrix degradation. The objective of this study was to determine the effect of female hormones on this initial cascade. Accordingly, an aortocaval fistula (Fist) was created in 7-wk-old Int and OVX rats, which, together with sham-operated (sham) controls, were studied at 1, 3, and 5 days postsurgery. In Int-Fist rats, myocardial mast cell density, collagen volume fraction, endothelin (ET)-1, stem cell factor (SCF), and TNF-α remained at control levels or were minimally elevated throughout the study period. This was not the case in the OVX-Fist group, where the initial response included significant increases in mast cell density, collagen degradation, ET-1, SCF, and TNF-α. These events in the OVX-Fist group were abolished by prefistula treatment with a mast cell stabilizer nedocromil. Of note was the observation that ET-1, TNF-α, SCF, and collagen volume fraction values for the OVX-sham group were greater than those of the Int-sham group, suggesting that the reduction of female hormones alone results in major myocardial changes. We concluded that female hormone-related cardioprotection to the volume stressed myocardium is the result of an altered mast cell phenotype and/or the prevention of mast cell activation.
Subject(s)
Cardiomegaly/physiopathology , Estrogens/metabolism , Heart Failure/physiopathology , Mast Cells/physiology , Ventricular Remodeling/physiology , Animals , Anti-Inflammatory Agents/pharmacology , Collagen/metabolism , Endothelin-1/metabolism , Female , Mast Cells/drug effects , Nedocromil/pharmacology , Ovariectomy , Phenotype , Rats , Rats, Sprague-Dawley , Stem Cell Factor/metabolism , Tumor Necrosis Factor-alpha/metabolism , Ventricular Remodeling/drug effectsABSTRACT
Mast cells are involved in immune disorders so that many of the proinflammatory and tissue destructive mediators produced by these cells have been implicated in the pathogenesis of rheumatoid arthritis. This scenario prompted us to investigate the correlation between mast cell degranulation and neutrophil influx within the digits and knees joints of arthritic mice assessing what could be the functional role(s) of joint mast cells in the response to collagen immunization. DBA/1J mice were submitted to collagen-induced arthritis and disease was assessed on day 21, 32 and 42 post-immunization. Pharmacological treatment with the glucocorticoid prednisolone, commonly used in the clinic, and nedocromil, a mast cell stabilizer, was performed from day 21 to 30. Arthritis develop after immunization, gradually increased up to day 42. Neutrophil infiltration peaked on day 32 and 21, in the digits and knees, respectively, showing an unequal pattern of recruitment between these tissues. This difference emerged for mast cells: they peaked in the digits on day 21, but a higher degree of degranulation could be measured in the knee joints. Uneven modulation of arthritis occurred after treatment of mice with prednisolone or nedocromil. Neutrophils migration to the tissue was reduced after both therapies, but only prednisolone augmented mast cell migration to the joints. Nedocromil exerted inhibitory properties both on mast cell proliferation and migration, more effectively on the digit joints. Thus, collagen induced an inflammatory process characterized by tissue mast cells activation and degranulation, suggesting a potential driving force in propagating inflammatory circuits yielding recruitment of neutrophils. However, the different degree of affected joint involvement suggests a time-related implication of digits and knees during collagen-induced arthritis development. These results provide evidence for local alterations whereby mast cells contribute to the initiation of inflammatory arthritis and may be targeted in intervention strategies.
Subject(s)
Arthritis, Experimental/immunology , Mast Cells/immunology , Neutrophil Infiltration/immunology , Neutrophils/immunology , Animals , Anti-Inflammatory Agents/pharmacology , Arthritis, Experimental/drug therapy , Arthritis, Experimental/pathology , Forelimb/drug effects , Forelimb/pathology , Glucocorticoids/pharmacology , Hindlimb/drug effects , Hindlimb/pathology , Knee Joint/drug effects , Knee Joint/immunology , Knee Joint/pathology , Leukocyte Count , Mast Cells/drug effects , Mast Cells/pathology , Mice , Mice, Inbred DBA , Nedocromil/pharmacology , Neutrophil Infiltration/drug effects , Neutrophils/drug effects , Neutrophils/pathology , Prednisolone/pharmacology , Synovial Membrane/drug effects , Synovial Membrane/immunology , Synovial Membrane/pathology , Time FactorsABSTRACT
OBJECTIVE: To determine whether the inhibitory action of the antiallergic cromone "mast cell stabilizing" drugs on polymorphonuclear leukocyte (PMN) trafficking is mediated through an annexin-A1 (Anx-A1) dependent mechanism. METHODS AND RESULTS: Intravital microscopy was used to monitor the actions of cromones in the inflamed microcirculation. Reperfusion injury provoked a dramatic increase in adherent and emigrated leukocytes in the mesenteric vascular bed, associated with augmented tissue levels of myeloperoxidase. Nedocromil, 2 to 20 mg/kg, significantly (P<0.05) inhibited cell adhesion and emigration, as well as myeloperoxidase release, in wild-type but not Anx-A1(-/-) mice. Short pretreatment of human PMNs with nedocromil, 10 nmol/L, inhibited cell adhesion (P<0.05) in the flow chamber assay, and this effect was reversed by specific anti-AnxA1 or a combination of antiformyl peptide receptors 1 and 2, but not irrelevant control, antibodies. Western blotting experiments revealed that cromones stimulate protein kinase C-dependent phosphorylation and release Anx-A1 in human PMNs. CONCLUSIONS: We propose a novel mechanism to explain the antiinflammatory actions of cromones on PMN trafficking, an effect that has long puzzled investigators.
Subject(s)
Annexin A1/metabolism , Anti-Allergic Agents/pharmacology , Anti-Inflammatory Agents/pharmacology , Cell Adhesion/drug effects , Cromolyn Sodium/pharmacology , Endothelial Cells/drug effects , Nedocromil/pharmacology , Neutrophils/drug effects , Animals , Annexin A1/deficiency , Annexin A1/genetics , Blotting, Western , Cells, Cultured , Disease Models, Animal , Dose-Response Relationship, Drug , Endothelial Cells/immunology , Endothelial Cells/metabolism , Humans , Leukocyte Rolling/drug effects , Male , Mesenteric Vascular Occlusion/drug therapy , Mesenteric Vascular Occlusion/immunology , Mesenteric Vascular Occlusion/metabolism , Mice , Mice, Inbred C57BL , Mice, Knockout , Microcirculation/drug effects , Microscopy, Video , Neutrophils/immunology , Neutrophils/metabolism , Peritonitis/drug therapy , Peritonitis/immunology , Peritonitis/metabolism , Peroxidase/metabolism , Phosphorylation , Protein Kinase C/metabolism , Protein Transport , Receptors, Formyl Peptide/drug effects , Receptors, Formyl Peptide/metabolism , Reperfusion Injury/drug therapy , Reperfusion Injury/immunology , Reperfusion Injury/metabolism , Time FactorsABSTRACT
We report that the asthma drugs cromolyn disodium and nedocromil sodium are potent G-protein-coupled receptor 35 (GPR35) agonists. We utilized calcium flux and inositol phosphate accumulation assays to examine the pharmacology of these asthma drugs on the human, mouse and rat GPR35. The compounds were more potent on the human GPR35 than on mouse and rat receptors. In contrast, zaprinast, a known GPR35 agonist, was more potent on mouse and rat GPR35 than the human ortholog. We show by quantitative PCR that GPR35 is expressed in human mast cells, human basophils and human eosinophils. We also demonstrate that GPR35 mRNA is upregulated upon challenge with IgE antibodies. We show that, unlike zaprinast, a potent phosphodiesterase 5 (PDE5) inhibitor, cromolyn disodium and nedocromil sodium lack inhibitory activity towards PDE5. These findings suggest that GPR35 may play an important role in mast cell biology and be a potential target for the treatment of asthma.
Subject(s)
Anti-Asthmatic Agents/pharmacology , Cromolyn Sodium/pharmacology , Nedocromil/pharmacology , Receptors, G-Protein-Coupled/agonists , Animals , Basophils/drug effects , Basophils/metabolism , Cricetinae , Drug Delivery Systems , Eosinophils/drug effects , Eosinophils/metabolism , Humans , Immunoglobulin E/pharmacology , In Vitro Techniques , Mast Cells/drug effects , Mast Cells/metabolism , Mice , Phosphodiesterase 5 Inhibitors/pharmacology , Purinones/pharmacology , RNA, Messenger/biosynthesis , Rats , Receptors, G-Protein-Coupled/biosynthesisABSTRACT
SCOPE OF THE REVIEW: Our knowledge of the multifunctional nature of airway smooth muscle (ASM) has expanded rapidly in the last decade, but the underlying molecular mechanisms and how current therapies for obstructive airway diseases, such as asthma and chronic obstructive pulmonary disease (COPD), affect these are still being elucidated. Our current knowledge has built on the pharmacology of human ASM contraction and relaxation established prior to that and which is reviewed in detail elsewhere in this issue. The advent of methods to isolate and culture ASM cells, especially human ASM cells, has made it possible to study how they may contribute to airway remodelling through their synthetic, proliferative, and migratory capacities. Now the underlying molecular mechanisms of ASM growth factor secretion, extracellular matrix (ECM) production, proliferation and migration, as well as contraction and relaxation, are being determined. A complex network of signalling pathways leading to gene transcription in ASM cells permits this functional plasticity in healthy and diseased airways. This review is an overview of the effects of current therapies, and some of those in development, on key signalling pathways and transcription factors involved in these ASM functions.
Subject(s)
Anti-Asthmatic Agents/pharmacology , Asthma/drug therapy , Muscle, Smooth/drug effects , Respiratory System/drug effects , Signal Transduction/drug effects , Transcriptional Activation/drug effects , Adrenergic beta-Agonists/pharmacology , Adrenergic beta-Agonists/therapeutic use , Animals , Anti-Asthmatic Agents/therapeutic use , Asthma/metabolism , Cholinergic Antagonists/pharmacology , Cholinergic Antagonists/therapeutic use , Cromolyn Sodium/pharmacology , Cromolyn Sodium/therapeutic use , Drug Therapy, Combination , Glucocorticoids/pharmacology , Glucocorticoids/therapeutic use , Humans , Leukotriene Antagonists/pharmacology , Leukotriene Antagonists/therapeutic use , Muscle, Smooth/metabolism , Muscle, Smooth/physiology , Nedocromil/pharmacology , Nedocromil/therapeutic use , Respiratory System/metabolism , Signal Transduction/physiology , Transcription Factors/metabolismABSTRACT
The present study aimed to clarify the role of mast cells in colitis with relapse induced in Wistar rats by trinitrobenzenosulphonic acid. Colitis induction increased the histamine concentration in the colon, which peaked on day 26. The number of mast cells, probably immature, was ten times higher on day 8. Different from animals infected with intestinal parasites, after colitis remission, mast cells do not migrate to the spleen, showing that mast cell proliferation presents different characteristics depending on the inflammation stimuli. Treatment with sulfasalazine, doxantrazole, quercetin, or nedocromil did not increase the histamine concentration or the mast cell number in the colon on day 26, thereby showing absence of degranulation of these cells. In conclusion, although mast cell proliferation is associated with colitis, these cells and their mediators appear to play no clear role in the colitis with relapses.
Subject(s)
Colitis/chemically induced , Colitis/pathology , Mast Cells/pathology , Trinitrobenzenesulfonic Acid/pharmacology , Animals , Colitis/metabolism , Colon/drug effects , Colon/metabolism , Colon/pathology , Gastrointestinal Agents/pharmacology , Histamine/metabolism , Male , Mast Cells/drug effects , Mast Cells/metabolism , Microscopy , Nedocromil/pharmacology , Rats , Rats, Wistar , Sulfasalazine/pharmacology , Thioxanthenes/pharmacology , Xanthones/pharmacologyABSTRACT
1.We investigated the role of Annexin (ANX)-A1 and its receptor, ALX/FPR2, in the regulation of mast cell degranulation produced by compound 48/80. 2.Both human cord-blood derived mast cells (CBDMCs) and murine bone marrow derived mast cells (BMDMCs) release phosphorylated ANX-A1 during treatment with glucocorticoids or the mast cell 'stabilising' drugs ketotifen and nedocromil. 3.Compound 48/80 also stimulated ANX-A1 phosphorylation and release and this was also potentiated by nedocromil. Anti-ANX-A1 neutralising monoclonal antibodies (Mabs) enhanced the release of pro-inflammatory mediators in response to compound 48/80. 4.Nedocromil and ketotifen potently inhibited the release of histamine, PGD2, tryptase and ß-hexosaminidase from mast cells challenged with compound 48/80. Anti-ANX-A1 neutralising Mabs prevented the inhibitory effect of these drugs. 5.BMDMCs derived from Anx-A1−/− mice were insensitive to the inhibitory effects of nedocromil or ketotifen but cells retained their sensitivity to the inhibitory action of hu-r-ANX-A1. 6.The fpr2/3 antagonist WRW4 blocked the action of nedocromil on PGD2, but not histamine, release. BMDMCs derived from fpr2/3−/− mice were insensitive to the inhibitory effects of nedocromil on PGD2, but not histamine release. 7.Compound 48/80 stimulated both p38 and JNK phosphorylation in CBDMCs and this was inhibited by nedocromil. Inhibition of p38 phosphorylation was ANX-A1 dependent. 8.We conclude that ANX-A1 is an important regulator of mast cell reactivity to compound 48/80 exerting a negative feedback effect through a mechanism that depends at least partly on the FPR receptor.
Subject(s)
Annexin A1/metabolism , Cell Degranulation/physiology , Mast Cells/drug effects , Receptors, Formyl Peptide/metabolism , Receptors, Lipoxin/metabolism , p-Methoxy-N-methylphenethylamine/pharmacology , Animals , Anti-Allergic Agents/pharmacology , Anti-Inflammatory Agents/pharmacology , Bone Marrow Cells/cytology , Dexamethasone/pharmacology , Fetal Blood/cytology , Humans , Indoles/pharmacology , Ketotifen/pharmacology , MAP Kinase Kinase 4/metabolism , Maleimides/pharmacology , Mast Cells/physiology , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Knockout , Nedocromil/pharmacology , Protein Kinase C/metabolism , Protein Kinase Inhibitors/pharmacology , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
Estrogen regulation of myocardial chymase and chymase effects on cardiac remodeling are unknown. To test the hypothesis that estrogen prevents pressure overload-induced adverse cardiac remodeling by inhibiting mast cell (MC) chymase release, transverse aortic constriction or sham surgery was performed in 7-week-old intact and ovariectomized (OVX) rats. Three days before creating the constriction, additional groups of OVX rats began receiving 17ß-estradiol, a chymase inhibitor, or a MC stabilizer. Left ventricular function, cardiomyocyte size, collagen volume fraction, MC density and degranulation, and myocardial and plasma chymase levels were assessed 18 days postsurgery. Aortic constriction resulted in ventricular hypertrophy in intact and OVX groups, whereas collagen volume fraction was increased only in OVX rats. Chymase protein content was increased by aortic constriction in the intact and OVX groups, with the magnitude of the increase being greater in OVX rats. MC density and degranulation, plasma chymase levels, and myocardial active transforming growth factor-ß1 levels were increased by aortic constriction only in OVX rats. Estrogen replacement markedly attenuated the constriction-increased myocardial chymase, MC density and degranulation, plasma chymase, and myocardial active transforming growth factor-ß1, as well as prevented ventricular hypertrophy and increased collagen volume fraction. Chymostatin attenuated the aortic constriction-induced ventricular hypertrophy and collagen volume fraction in the OVX rats similar to that achieved by estrogen replacement. Nedocromil yielded similar effects, except for the reduction of chymase content. We conclude that the estrogen-inhibited release of MC chymase is responsible for the cardioprotection against transverse aortic constriction-induced adverse cardiac remodeling.
Subject(s)
Chymases/metabolism , Estradiol/pharmacology , Hypertrophy, Left Ventricular/prevention & control , Mast Cells/drug effects , Ventricular Remodeling/drug effects , Animals , Cardiomyopathy, Hypertrophic/complications , Cell Degranulation/drug effects , Cell Size/drug effects , Collagen/analysis , Estrogen Replacement Therapy , Female , Hypertrophy, Left Ventricular/etiology , Mast Cells/enzymology , Mast Cells/metabolism , Mast Cells/physiology , Myocardium/enzymology , Myocytes, Cardiac/drug effects , Myocytes, Cardiac/enzymology , Myocytes, Cardiac/ultrastructure , Nedocromil/pharmacology , Oligopeptides/pharmacology , Organ Size/drug effects , Ovariectomy , Random Allocation , Rats , Rats, Sprague-Dawley , Transforming Growth Factor beta1/metabolism , Ventricular Dysfunction, Left/etiology , Ventricular Dysfunction, Left/prevention & controlABSTRACT
The extent of mast cell direct involvement in fibrosis is not defined as yet. In the present study we assessed whether long-term co-culture (up to 7 d) of functionally active rat peritoneal mast cells with 3T3 mouse fibroblasts and mass cell activation can affect fibroblast proliferation and collagen production. Co-culture of subconfluent 3T3 fibroblasts with resting mast cells or with mast cells stimulated by alpha IgE (1:35) or repeatedly activated by low concentrations of compound 48/80 (0.25-0.75 microgram/ml) did not alter fibroblast proliferation. However, fibroblast proliferation was increased significantly (100-130%) when mast cells were repeatedly activated with higher concentrations of compound 48/80 (1-3 micrograms/ml). Repeated mast cell activation by compound 48/80 (0.25 microgram/ml) caused a twofold increase in collagen production and this was reduced by 63% by the mast cell stabilizer nedocromil sodium (10(-5) M). At the same time, co-culture of 3T3 fibroblasts with unstimulated or immunologically activated mast cells did not modulate their collagen production. In conclusion, we have demonstrated that mast cell activation, under certain conditions, can enhance significantly 3T3 fibroblast proliferation and collagen production, thus indicating a direct mast cell involvement in the fibrotic processes.
Subject(s)
3T3 Cells/pathology , Mast Cells/physiology , Animals , Cell Division/physiology , Collagen/biosynthesis , Fibroblasts/metabolism , Fibrosis/etiology , Mice , Nedocromil/pharmacology , p-Methoxy-N-methylphenethylamine/pharmacologyABSTRACT
Metachromatic cells are increased in the airways of asthmatic patients. We obtained metachromatic cells from asthmatic airways using an induced sputum technique. The histamine release following ConA, anti-IgE and anti-IgE + IL3 from those cells were evaluated before and following the addition of cromoline Na and nedocromil Na. Metachromatic cells had a higher rate of spontaneous histamine release when compared to peripheral basophils. Cromoline Na and nedocromil Na inhibited histamine release from triggered metachromatic cells but not from peripheral basophils. It is concluded that airway metachromatic cells from asthmatics behave differently than peripheral basophils.
Subject(s)
Anti-Asthmatic Agents/pharmacology , Asthma/immunology , Basophils/drug effects , Cromolyn Sodium/pharmacology , Histamine Release/drug effects , Nedocromil/pharmacology , Adult , Antibodies, Anti-Idiotypic/pharmacology , Asthma/blood , Bronchi/cytology , Concanavalin A/pharmacology , Female , Forced Expiratory Volume , Humans , Immunoglobulin E/immunology , Interleukin-3/pharmacology , Male , Sputum/cytologyABSTRACT
1. Platelet activating factor (PAF), leukotriene B4 (LTB4) and interleukin-5 (IL-5) are potent chemoattractants for guinea-pig eosinophils, which may be involved in eosinophil recruitment and up-regulation in allergic diseases. Eosinophils from the bronchoalveolar lavage fluid (BALF) of ovalbumin-sensitized guinea-pigs were collected 24 h after antigen provocation and migration induced by PAF, LTB4 and rhIL-5 was studied. 2. Total BALF content and distribution of eosinophils were greater in immunized, ovalbumin-challenged guinea-pigs (5.0 +/- 0.8 x 10(6)/guinea-pig; 12 +/- 1%) than in immunized, saline-challenged animals (3.0 +/- 0.7 x 10(6)/guinea-pig; 7 +/- 1%). 3. The chemoattraction of eosinophils isolated on a metrizamide gradient was studied in micro-Boyden chambers, results being expressed as the number of migrating cells (mean +/- s.e. mean). PAF and LTB4-induced migration of eosinophils from immunized and OA-challenged guinea-pigs were significantly enhanced, as compared to immunized and saline-challenged animals (170 +/- 36 vs 35 +/- 9 migrating eosinophils for 10 nM PAF; 271 +/- 60 vs 110 +/- 19 for 1 nM LTB4). 4. The IL-5 antibody TRFK-5, in vivo, reduced eosinophil recruitment in BALF of antigen-challenged immunized animals as well as the enhanced responsiveness of eosinophils from the challenged animals, suggesting a role for IL-5 in the priming of eosinophils in vivo. 5. In contrast to TRFK-5, nedocromil sodium reduced to a similar extent eosinophil, macrophage and lymphocyte recruitment into the BALF of antigen-challenged, but failed to down-regulate the enhanced responsiveness of eosinophils from the challenged animals. 6. The increased eosinophil content in lungs from antigen-challenged guinea-pigs is thus selectively reduced by the anti-IL-5 antibody, which also attenuates the concomitant enhancement of the eosinophil responsiveness, supporting the concept that IL-5 is essential for recruitment and priming of eosinophils in vivo. In contrast, nedocromil sodium reduced non-selectively the total cell recruitment to the airways,but failed to attenuate the enhanced responsiveness of those eosinophils which migrated, indicating that its effects involve a different target.
Subject(s)
Antigens/immunology , Bronchoalveolar Lavage Fluid/cytology , Eosinophils/physiology , Interleukin-5/physiology , Animals , Antibodies, Monoclonal/immunology , Cell Movement , Guinea Pigs , Immunization , Leukotriene B4/pharmacology , Male , Nedocromil/pharmacology , Ovalbumin/immunology , Platelet Activating Factor/pharmacologyABSTRACT
1. Sodium metabisulphite (MBS) can induce bronchoconstriction in patients with asthma. We investigated the effects of MBS aerosol on bronchial blood velocity (Vbr) and pulmonary resistance in intubated conscious sheep. 2. Bronchial blood velocity was measured by implanting a 20 MHz ultrasonic Doppler flow probe on the common bronchial branch of the bronchoesophageal artery. 3. Inhaled MBS induced a dose-dependent, transient increase in Vbr lasting for a few minutes without any changes in aortic and pulmonary artery pressures. There was some tachyphylaxis of the Vbr response to successive inhalations of MBS. 4. The cholinoceptor antagonist, ipratropium bromide and the H1 and H2 histamine antagonists, chlorpheniramine and cimetidine, had no significant effect on MBS-induced increase on Vbr. The loop diuretic, frusemide, and the anti-inflammatory drug, nedocromil sodium, which both inhibit MBS-induced bronchoconstriction in patients with asthma, were also without effect. 5. We conclude that MBS induces bronchial vasodilatation in conscious sheep, and that this effect is not dependent on the release of histamine or other mediators, or an activation of cholinergic pathways.
Subject(s)
Bronchi/blood supply , Pulmonary Circulation/drug effects , Sulfites/pharmacology , Administration, Inhalation , Aerosols , Animals , Cholinergic Antagonists , Consciousness , Dose-Response Relationship, Drug , Furosemide/pharmacology , Histamine H1 Antagonists/pharmacology , Histamine H2 Antagonists/pharmacology , Nedocromil/pharmacology , Sheep , Sulfites/pharmacokinetics , Tachyphylaxis , Vascular Resistance/drug effectsABSTRACT
Chromones (sodium cromoglycate and sodium nedocromil) block cell swelling-activated Cl- channels in NIH-3T3 fibroblasts and endothelial cells. This has led to hypothesize that cell volume regulation might be involved in asthma pathogenesis. Using whole-cell patch-clamp experiments, we studied the effect of chromones on volume-sensitive Cl- currents in transformed human tracheal epithelial cells (9HTEo-) and in primary cultures of human bronchial epithelial cells (BE). Cl- currents activated by hypotonic shock were poorly blocked by extracellular nedocromil or cromoglycate. The block was voltage-dependent since it was observed only at positive membrane potentials. At the concentration of 5 mM, the current inhibition by both chromones at +80 mV was about 40% for 9HTEo- and only 20% for BE. Intracellular application of chromones elicited a voltage-independent inhibition in 9HTEo- cells. Under this condition, volume-sensitive Cl- currents were reduced at all membrane potentials (60 and 45% inhibition by 2 mM nedocromil and cromoglycate respectively). In contrast intracellular chromones were ineffective in BE cells. The relative refractoriness to chromones, in contrast with the high sensitivity shown by other Cl- channels, suggests that the epithelial volume-sensitive Cl- channel is not involved in asthma.
Subject(s)
Anti-Asthmatic Agents/pharmacology , Chloride Channels/antagonists & inhibitors , Chlorides/physiology , Cromolyn Sodium/pharmacology , Nedocromil/pharmacology , Trachea/drug effects , 3T3 Cells/drug effects , 3T3 Cells/physiology , Animals , Cells, Cultured , Chloride Channels/physiology , Epithelial Cells/drug effects , Epithelial Cells/physiology , Humans , Kinetics , Membrane Potentials/drug effects , Membrane Potentials/physiology , Mice , Sensitivity and Specificity , Trachea/physiologyABSTRACT
1. We have investigated the novel naturally occurring marine compound, IZP-94005 (contignasterol), as a potential anti-asthma agent, using both in vivo and in vitro models of allergen-induced bronchoconstriction and airway smooth muscle contraction. 2. Tracheal rings from ovalbumin (OA)-sensitized guinea-pigs were treated with various concentrations of IZP-94005 for 20 min prior to challenge with ovalbumin. IZP-94005 (3-30 microM) inhibited responses of sensitized tracheal rings stimulated with OA in a concentration-dependent manner, with an IC50 of 10 microM. 3. IZP-94005 (10 microM) had no effect on carbachol-induced contractions of sensitized guinea-pig tracheal rings, although it did inhibit histamine-induced responses of OA sensitized guinea-pig tracheal rings. 4. The effects of IZP-94005 in vivo were examined using OA-sensitized guinea-pigs which were tracheotomized under anaesthesia and placed in a body plethysmograph. Measurements of lung resistance and compliance were performed by isovolumetric analysis of volume and trans-pulmonary pressure. 5. IZP-94005 (50 and 200 micrograms kg-1), by inhalation 20 min prior to OA challenge caused significant inhibition of the increase in lung resistance induced by OA in sensitized guinea-pigs, compared to vehicle-treated animals. Nedocromil sodium (20 mg kg-1), with a similar protocol, also inhibited OA-induced responses in this model. 6. We therefore suggest that IZP-94005 is a good candidate for further investigation as a possible antiasthma agent.
Subject(s)
Anti-Asthmatic Agents/therapeutic use , Bronchial Hyperreactivity/drug therapy , Bronchoconstriction/drug effects , Muscle, Smooth/drug effects , Sterols/therapeutic use , Administration, Inhalation , Airway Resistance/drug effects , Analysis of Variance , Animals , Anti-Asthmatic Agents/administration & dosage , Anti-Asthmatic Agents/pharmacology , Bronchial Hyperreactivity/chemically induced , Carbachol/pharmacology , Guinea Pigs , Histamine Release/drug effects , Lung/drug effects , Lung/metabolism , Muscle Contraction/drug effects , Nedocromil/administration & dosage , Nedocromil/pharmacology , Nedocromil/therapeutic use , Ovalbumin/administration & dosage , Plethysmography , Sterols/administration & dosage , Sterols/pharmacology , Trachea/drug effectsABSTRACT
OBJECTIVE: Nedocromil sodium (nedocromil) improves the clinical condition of asthmatic subjects but its mechanism of action is not fully understood. This study aimed to determine whether nedocromil alters the ability of contractile and relaxant non-adrenergic, non-cholinergic neural (NANC) responses to stabilise tone by inhibiting or potentiating these responses in bronchial smooth muscle and, if so, whether the action is on a pre- or postjunctional level. RESULTS: Nedocromil attenuated contractile but not relaxant NANC responses (elicited by electric field stimulation) significantly (P < 0.05) in guinea pig main bronchi in vitro. However, the ability of NANC responses to stabilise tone (convergence effect) was not significantly impaired by nedocromil. Furthermore, nedocromil did not significantly shift the concentration response curve (-log EC50) to neurokinin A (NKA), the dominating contractile NANC transmitter, or alter the maximum response to NKA (P > 0.05). Submaximum or maximum contractile responses to histamine were not markedly affected by nedocromil (P > 0.05). CONCLUSIONS: Nedocromil exerts selective neural inhibition of the contractile but not of the relaxant NANC responses on a pre-junctional level in bronchial smooth muscle. Nedocromil does not, however, markedly impair the ability of NANC response to stabilise bronchial smooth muscle tone.
Subject(s)
Anti-Asthmatic Agents/pharmacology , Bronchi/drug effects , Bronchi/physiology , Muscle Contraction/drug effects , Nedocromil/pharmacology , Animals , Bronchial Provocation Tests , Female , Guinea Pigs , Histamine , In Vitro Techniques , Muscle Relaxation/drug effects , Muscle Relaxation/physiology , Muscle, Smooth/drug effects , Muscle, Smooth/physiology , Neurokinin A/pharmacology , Neurons/drug effects , Neurons/physiology , Receptors, Adrenergic/drug effects , Receptors, Cholinergic/drug effectsABSTRACT
Nedocromil sodium is commonly suggested to reduce allergic inflammation by inhibiting mediator release from mast cells. However, nedocromil also exhibits a wide range of additional anti-inflammatory activities, including inhibition of increased vascular permeability induced by individual mediators such as histamine. In the present study, we have further characterized the mode of action of nedocromil in a rat model for hind paw edema. Mast cell-dependent edema was induced with compound 48/80 (edema response mainly due to 5-hydroxytryptamine release), and direct mediator-induced plasma extravasation was evoked by exogenous 5-hydroxytryptamine (both agents injected locally). Local pretreatment with nedocromil for 20 min dose-dependently inhibited the edema evoked by compound 48/80 more effectively than that induced by 5-hydroxytryptamine. However, after 2 h pretreatment, both the 5-hydroxytryptamine-and compound 48/80-induced edema responses were inhibited to approximately the same extent by a range of concentrations of nedocromil, as well as by dexamethasone. Local inhibition of RNA/protein synthesis with actinomycin-D abolished the effects of both dexamethasone and nedocromil (2 h local pretreatment). We thus conclude that nedocromil can produce an 'anti-exudative' effect that is independent of inhibition of mast cell mediator release, is slow in onset, and requires de novo protein synthesis.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Dexamethasone/pharmacology , Glucocorticoids/pharmacology , Nedocromil/pharmacology , Animals , Dose-Response Relationship, Drug , Edema/chemically induced , Edema/drug therapy , Male , Mast Cells/drug effects , Rats , Rats, Sprague-Dawley , Serotonin Antagonists/pharmacologyABSTRACT
In the rat trachea, two types of mast cells have been identified, connective tissue mast cells and mucosal mast cells. Their different characteristics may account for their different biological functions. The role of connective tissue mast cells in tracheal contraction as one feature of the immediate reaction of asthma was studied in vitro in isolated trachea, using tissue derived from mast cell-deficient (Ws/Ws) rats, heterozygous (Ws/+) rats and control (+/+) rats, and compound 48/80 as a potent inducer of mast cell degranulation. The contractile response of tracheas from the three types of rats was also studied upon exposure to the following spasmogens: histamine, 5-hydroxytryptamine (5-HT), and carbachol. Histamine content in tissues reflected the differing mast cell numbers in strips from the three rat types. It was found that carbachol and 5-HT elicited tracheal contraction in a similar manner in strips from the three types of rats. Histamine had no contractile effect. Compound 48/80, at a dose of 25 microg/ml, elicited contraction in tracheas from both control (+/+) and heterozygous (Ws/+), but not in trachea from Ws/Ws rats. Compound 48/80-induced contractions in tracheas from +/+ rats were inhibited by 0.1 microM ketanserin and 0.1 microM nedocromil, but not by 0.1 microM mepyramine. Enzyme histochemistry confirmed that the degranulation occurred in connective tissue mast cells, but not in mucosal mast cells. We concluded that connective tissue mast cells play an important role in rat tracheal contraction via 5-HT release induced by compound 48/80. In addition, the specific mast cell-deficient (Ws/Ws) rats provide a good tool for studying the roles of mast cells in airway system.