ABSTRACT
Increased insulin is associated with obesity-related airway hyperreactivity and asthma. We tested whether the use of metformin, an antidiabetic drug used to reduce insulin resistance, can reduce circulating insulin, thereby preventing airway hyperreactivity in rats with dietary obesity. Male and female rats were fed a high- or low-fat diet for 5 wk. Some male rats were simultaneously treated with metformin (100 mg/kg orally). In separate experiments, after 5 wk of a high-fat diet, some rats were switched to a low-fat diet, whereas others continued a high-fat diet for an additional 5 wk. Bronchoconstriction and bradycardia in response to bilateral electrical vagus nerve stimulation or to inhaled methacholine were measured in anesthetized and vagotomized rats. Body weight, body fat, caloric intake, fasting glucose, and insulin were measured. Vagally induced bronchoconstriction was potentiated only in male rats on a high-fat diet. Males gained more body weight, body fat, and had increased levels of fasting insulin compared with females. Metformin prevented development of vagally induced airway hyperreactivity in male rats on high-fat diet, in addition to inhibiting weight gain, fat gain, and increased insulin. In contrast, switching rats to a low-fat diet for 5 wk reduced body weight and body fat, but it did not reverse fasting glucose, fasting insulin, or potentiation of vagally induced airway hyperreactivity. These data suggest that medications that target insulin may be effective treatment for obesity-related asthma.
Subject(s)
Asthma/drug therapy , Bronchial Hyperreactivity/drug therapy , Bronchoconstriction , Diet, High-Fat/adverse effects , Hyperinsulinism/prevention & control , Metformin/pharmacology , Obesity/complications , Animals , Asthma/chemically induced , Asthma/metabolism , Asthma/pathology , Bronchial Hyperreactivity/chemically induced , Bronchial Hyperreactivity/metabolism , Bronchial Hyperreactivity/pathology , Bronchoconstrictor Agents/toxicity , Female , Glucose/metabolism , Hyperinsulinism/etiology , Hyperinsulinism/metabolism , Hyperinsulinism/pathology , Hypoglycemic Agents/pharmacology , Male , Methacholine Chloride/toxicity , Rats , Rats, Sprague-Dawley , Vagus Nerve/drug effects , Weight GainABSTRACT
OBJECTIVE: Immediately preceding sudden unexpected death in epilepsy (SUDEP), patients experienced a final generalized tonic-clonic seizure (GTCS), rapid ventilation, apnea, bradycardia, terminal apnea, and asystole. Whether a progressive pathophysiology develops and increases risk of SUDEP remains unknown. Here, we determined (a) heart rate, respiratory rate, and blood oxygen saturation (SaO2 ) in low-risk and high-risk knockout (KO) mice; and (b) whether blocking receptors for orexin, a cardiorespiratory neuromodulator, influences cardiorespiratory function mice or longevity in high-risk KO mice. METHODS: Heart rate and SaO2 were determined noninvasively with ECGenie and pulse oximetry. Respiration was determined with noninvasive airway mechanics technology. The role of orexin was determined within subject following acute treatment with a dual orexin receptor antagonist (DORA, 100 mg/kg). The number of orexin neurons in the lateral hypothalamus was determined with immunohistochemistry. RESULTS: Intermittent bradycardia was more prevalent in high-risk KO mice, an effect that may be the result of increased parasympathetic drive. High-risk KO mice had more orexin neurons in the lateral hypothalamus. Blocking of orexin receptors differentially influenced heart rate in KO, but not wild-type (WT) mice. When DORA administration increased heart rate, it also decreased heart rate variability, breathing frequency, and/or hypopnea-apnea. Blocking orexin receptors prevented the methacholine (MCh)-induced increase in breathing frequency in KO mice and reduced MCh-induced seizures, via a direct or indirect mechanism. DORA improved oxygen saturation in KO mice with intermittent hypoxia. Daily administration of DORA to high-risk KO mice increased longevity. SIGNIFICANCE: High-risk KO mice have a unique cardiorespiratory phenotype that is characterized by progressive changes in five interdependent endpoints. Blocking of orexin receptors attenuates some of these endpoints and increases longevity, supporting the notion that windows of opportunity for intervention exist in this preclinical SUDEP model.
Subject(s)
Apnea/genetics , Bradycardia/genetics , Epilepsy/genetics , Hypoxia/genetics , Kv1.1 Potassium Channel/genetics , Sudden Unexpected Death in Epilepsy , Animals , Apnea/physiopathology , Bradycardia/physiopathology , Epilepsy/physiopathology , Heart Rate/drug effects , Heart Rate/physiology , Hypothalamic Area, Lateral/metabolism , Hypothalamic Area, Lateral/pathology , Hypoxia/physiopathology , Methacholine Chloride/toxicity , Mice , Mice, Knockout , Neurons/metabolism , Neurons/pathology , Orexin Receptor Antagonists/pharmacology , Orexins/metabolism , Oximetry , Oxygen , Parasympathetic Nervous System/physiopathology , Parasympathomimetics/toxicity , Respiratory Rate/drug effects , Seizures/chemically inducedABSTRACT
Asthma is a chronic inflammatory lung disease of the airway; the incidence and prevalence of asthma remain high worldwide. Astragaloside IV (AS-IV) is the main active constituent of Astragalus membranaceus. Accumulating evidence suggests that AS-IV possesses anti-inflammatory and anti-asthmatic ability, but the potential molecular mechanism is required to further clarify. In this study, the anti-asthmatic effects of AS-IV on mice with ovalbumin (OVA)-induced allergic inflammation were analysed. We analysed airway hyperresponsiveness (AHR), numbers of inflammatory cells, inflammation situation in lung tissue and cytokines level in bronchoalveolar lavage fluid (BALF) between OVA-induced mice with and without AS-IV treatment. Moreover, we explored the possible signalling pathway behind the anti-asthmatic effects. Our results revealed that AS-IV treatment ameliorates airway inflammation and AHR in an OVA-induced asthma model. Besides, AS-IV treatment inhibits the interleukin (IL)-4, -5 and -13 production, and further study indicated that AS-IV treatment downregulates the expression level of p-JAK2/p-STAT6 proteins. Taken together, the present study suggested that the inhibitory effects of AS-IV on asthma therapy are at least partially involved in inhibiting the JAK2/STAT6 signalling pathway.
Subject(s)
Asthma/chemically induced , Gene Expression Regulation/drug effects , Janus Kinase 2/metabolism , STAT6 Transcription Factor/metabolism , Saponins/pharmacology , Signal Transduction/drug effects , Triterpenes/pharmacology , Animals , Asthma/drug therapy , Bronchoalveolar Lavage Fluid/cytology , Female , Janus Kinase 2/genetics , Leukocytes/physiology , Male , Methacholine Chloride/toxicity , Mice , Mice, Inbred BALB C , Ovalbumin/toxicity , Parasympathomimetics/toxicity , STAT6 Transcription Factor/geneticsABSTRACT
BACKGROUND: Ultra-long-acting ß2 agonists (uLABA) are relatively new anti-asthma medications of which there are three different formulations currently available: olodaterol, indacaterol, and vilanterol. The first 2 formulations have been shown to exert bronchoprotective effects; they are able to prevent airway smooth muscle contraction on exposure to constricting stimuli. However, studies have found that these 2 drugs produce different degrees and durations of bronchoprotection against methacholine. OBJECTIVE: The objective of this study was to investigate the degree of bronchoprotection provided by vilanterol against methacholine-induced bronchoconstriction. METHODS: Fourteen patients with mild-to-moderate asthma (8 male; baseline percent predicted forced expiratory volume in 1 second [FEV1] > 65%; provocative concentration of methacholine causing a 20% reduction in FEV1 [PC20] ≤ 8 mg/mL) completed this randomized, double-blind, 3-way crossover study. Methacholine challenges were performed before treatment administration (placebo, 100 µg fluticasone furoate, or 25 µg vilanterolâ¯+â¯100 µg fluticasone furoate) and at 0.5 and 24 hours posttreatment. Each treatment arm was separated by a minimum 7-day washout period. A combination therapy of vilanterol+fluticasone furoate was used, because vilanterol is not available as a monotherapy. RESULTS: Significant bronchoprotection was evident after the combination treatment at both 0.5 and 24 hours with doubling dose shifts in methacholine PC20 of 2.0 (Pâ¯=â¯.0004) and 1.6 (Pâ¯=â¯.0001), respectively. Clinically significant bronchodilation was only recorded at 24 hours after combination treatment (P < .05). CONCLUSION: These findings suggest that vilanterol (in combination with fluticasone furoate) provides significant bronchoprotection against methacholine-induced bronchoconstriction for at least 24 hours in patients with mild-to-moderate asthma. CLINICAL TRIAL REGISTRATION: clinicaltrials.gov (NCT03315000).
Subject(s)
Adrenergic beta-2 Receptor Agonists/therapeutic use , Androstadienes/therapeutic use , Asthma/drug therapy , Benzyl Alcohols/therapeutic use , Bronchoconstriction/drug effects , Bronchodilator Agents/therapeutic use , Chlorobenzenes/therapeutic use , Cross-Over Studies , Female , Forced Expiratory Volume/drug effects , Humans , Male , Methacholine Chloride/toxicityABSTRACT
Airway hyperresponsiveness often constitutes a primary outcome in respiratory studies in mice. The procedure commonly employs aerosolized challenges, and results are typically reported in terms of bronchoconstrictor concentrations loaded into the nebulizer. Yet, because protocols frequently differ across studies, especially in terms of aerosol generation and delivery, direct study comparisons are difficult. We hypothesized that protocol variations could lead to differences in aerosol delivery efficiency and, consequently, in the dose delivered to the subject, as well as in the response. Thirteen nebulization patterns containing common protocol variations (nebulization time, duty cycle, particle size spectrum, air humidity, and/or ventilation profile) and using increasing concentrations of methacholine and broadband forced oscillations (flexiVent, SCIREQ, Montreal, Qc, Canada) were created, characterized, and studied in anesthetized naïve A/J mice. A delivered dose estimate calculated from nebulizer-, ventilator-, and subject-specific characteristics was introduced and used to account for protocol variations. Results showed that nebulization protocol variations significantly affected the fraction of aerosol reaching the subject site and the delivered dose, as well as methacholine reactivity and sensitivity in mice. From the protocol variants studied, addition of a slow deep ventilation profile during nebulization was identified as a key factor for optimization of the technique. The study also highlighted sensitivity differences within the lung, as well as the possibility that airway responses could be selectively enhanced by adequate control of nebulizer and ventilator settings. Reporting results in terms of delivered doses represents an important standardizing element for assessment of airway hyperresponsiveness in mice.
Subject(s)
Methacholine Chloride/toxicity , Respiratory Hypersensitivity/chemically induced , Administration, Inhalation , Aerosols , Animals , Disease Models, Animal , Humans , Male , Mice , Nebulizers and Vaporizers/standards , Reference Standards , Research DesignABSTRACT
Chlorine (Cl2) inhalation induces severe oxidative lung injury and airway hyperresponsiveness (AHR) that lead to asthmalike symptoms. When inhaled, Cl2 reacts with epithelial lining fluid, forming by-products that damage hyaluronan, a constituent of the extracellular matrix, causing the release of low-molecular-weight fragments (L-HA, <300 kDa), which initiate a series of proinflammatory events. Cl2 (400 ppm, 30 min) exposure to mice caused an increase of L-HA and its binding partner, inter-α-trypsin-inhibitor (IαI), in the bronchoalveolar lavage fluid. Airway resistance following methacholine challenge was increased 24 h post-Cl2 exposure. Intratracheal administration of high-molecular-weight hyaluronan (H-HA) or an antibody against IαI post-Cl2 exposure decreased AHR. Exposure of human airway smooth muscle (HASM) cells to Cl2 (100 ppm, 10 min) or incubation with Cl2-exposed H-HA (which fragments it to L-HA) increased membrane potential depolarization, intracellular Ca(2+), and RhoA activation. Inhibition of RhoA, chelation of intracellular Ca(2+), blockade of cation channels, as well as postexposure addition of H-HA, reversed membrane depolarization in HASM cells. We propose a paradigm in which oxidative lung injury generates reactive species and L-HA that activates RhoA and Ca(2+) channels of airway smooth muscle cells, increasing their contractility and thus causing AHR.
Subject(s)
Asthma/drug therapy , Bronchial Hyperreactivity/drug therapy , Hyaluronic Acid/therapeutic use , Lung Injury/drug therapy , Oxidative Stress/drug effects , Alpha-Globulins/antagonists & inhibitors , Alpha-Globulins/biosynthesis , Alpha-Globulins/immunology , Animals , Bronchial Hyperreactivity/immunology , Bronchial Provocation Tests , Bronchoalveolar Lavage Fluid/cytology , Calcium/metabolism , Calcium Channel Blockers , Calcium Channels/metabolism , Cells, Cultured , Chlorine/toxicity , Enzyme Activation , Extracellular Matrix , Inflammation , Membrane Potentials/drug effects , Methacholine Chloride/toxicity , Mice , Mice, Inbred C57BL , Muscle Contraction/drug effects , Myocytes, Smooth Muscle , Patch-Clamp Techniques , Reactive Oxygen Species/metabolism , Trachea/metabolism , rho GTP-Binding Proteins/metabolism , rhoA GTP-Binding ProteinABSTRACT
Asthma is a chronic inflammatory disease of the small airways, with airway hyperresponsiveness (AHR) and inflammation as hallmarks. Recent studies suggest a role for arginase in asthma pathogenesis, possibly because arginine is the substrate for both arginase and NO synthase and because NO modulates bronchial tone and inflammation. Our objective was to investigate the importance of increased pulmonary arginase 1 expression on methacholine-induced AHR and lung inflammation in a mouse model of allergic asthma. Arginase 1 expression in the lung was ablated by crossing Arg1(fl/fl) with Tie2Cre(tg/-) mice. Mice were sensitized and then challenged with ovalbumin. Lung function was measured with the Flexivent. Adaptive changes in gene expression, chemokine and cytokine secretion, and lung histology were quantified with quantitative PCR, ELISA, and immunohistochemistry. Arg1 deficiency did not affect the allergic response in lungs and large-airway resistance, but it improved peripheral lung function (tissue elastance and resistance) and attenuated adaptive increases in mRNA expression of arginine-catabolizing enzymes Arg2 and Nos2, arginine transporters Slc7a1 and Slc7a7, chemokines Ccl2 and Ccl11, cytokines Tnfa and Ifng, mucus-associated epithelial markers Clca3 and Muc5ac, and lung content of IL-13 and CCL11. However, expression of Il4, Il5, Il10, and Il13 mRNA; lung content of IL-4, IL-5, IL-10, TNF-α, and IFN-γ protein; and lung pathology were not affected. Correlation analysis showed that Arg1 ablation disturbed the coordinated pulmonary response to ovalbumin challenges, suggesting arginine (metabolite) dependence of this response. Arg1 ablation in the lung improved peripheral lung function and affected arginine metabolism but had little effect on airway inflammation.
Subject(s)
Arginase/physiology , Asthma/physiopathology , Bronchial Hyperreactivity/pathology , Hypersensitivity/pathology , Lung/physiology , Pneumonia/pathology , Respiratory System/pathology , Airway Resistance/physiology , Animals , Blotting, Western , Bronchial Hyperreactivity/chemically induced , Bronchial Hyperreactivity/metabolism , Bronchoconstrictor Agents/toxicity , Chemokines/metabolism , Cytokines/metabolism , Dendritic Cells/cytology , Dendritic Cells/metabolism , Female , Gene Expression Profiling , Hypersensitivity/metabolism , Immunoenzyme Techniques , Lung/cytology , Macrophages/cytology , Macrophages/metabolism , Male , Methacholine Chloride/toxicity , Mice , Mice, Inbred C57BL , Mice, Knockout , Myeloid Cells/cytology , Myeloid Cells/metabolism , Ovalbumin/physiology , Pneumonia/chemically induced , Pneumonia/metabolism , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction , Respiratory System/drug effects , Respiratory System/metabolism , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Probiotics are normal inhabitants of the gastrointestinal tract of man and are widely considered to exert a number of beneficial effects in many diseases. But the mechanism by which they modulate the immune system is poorly understood. The present study was planned to explore the anti-allergic effect of Lactobacillus gasseri on a mouse model of allergic asthma. Dermatophoides pteronyssinus (Der p) sensitised and challenged BALB/c mice were orally administered via oral administration with three different doses of L. gasseri (low, 1 × 10(6) colony-forming units (CFU); medium, 2 × 10(6) CFU; high, 4 × 10(6) CFU), in 700 µl of PBS daily, starting from 2 weeks before Der p sensitisation for 4 weeks. After the allergen challenge, airway responsiveness to methacholine, influx of inflammatory cells to the lung, and cytokine levels in bronchoalveolar lavage (BAL) fluids and splenocytes culture were assessed. Our results showed that oral administration of a high dose of L. gasseri (4 × 10(6) CFU) decreased airway responsiveness to methacholine, attenuated the influx of inflammatory cells to the airways and reduced the levels of TNF-α, thymus and activation-regulated chemokine (TARC) and IL-17A in BAL fluids of Der p-sensitised and -challenged mice. Moreover, L. gasseri decreased IL-17A production in transforming growth factor-α and IL-6 stimulated splenocytes and cell numbers of IL-17 producing alveolar macrophages in L. gasseri-treated mice as compared to non-treated, Der p-sensitised and -challenged mice. In conclusion, oral administration with L. gasseri can attenuate major characteristics of allergen-induced airway inflammation and IL-17 pro-inflammatory immune response in a mouse model of allergic asthma, which may have clinical implication in the preventive or therapeutic potential in allergic asthma.
Subject(s)
Asthma/metabolism , Asthma/microbiology , Inflammation/prevention & control , Lactobacillus/classification , Probiotics , Th17 Cells/microbiology , Animals , Antibodies/blood , Antigens, Dermatophagoides/immunology , Asthma/immunology , Cytokines/genetics , Cytokines/metabolism , Enzyme-Linked Immunosorbent Assay , Female , Gene Expression Regulation , Immunoglobulin E , Immunoglobulin G/blood , Immunoglobulin G/classification , Inflammation/immunology , Lactobacillus/physiology , Methacholine Chloride/toxicity , Mice , Mice, Inbred BALB C , Th17 Cells/physiologyABSTRACT
Penh is a dimensionless index normally used to evaluate changes in the shape of the airflow pattern entering and leaving a whole-body flow plethysmograph as an animal breathes. The index is sensitive to changes in the distribution of area under the waveform during exhalation and increases in a nonlinear fashion as the normalized area increases near the beginning of the curve. Enhanced pause (Penh) has been used to evaluate changes in pulmonary function and as a method to evaluate airway reactivity. However, the use of Penh to assess pulmonary function has been challenged (Bates et al., 2004; Lundblad et al., 2002; Mitzner et al., 2003; Mitzner & Tankersley, 1998; Petak et al., 2001; Sly et al., 2005). The objective of this study was to show how Penh of the thorax and plethysmograph flow patterns are related. That relationship is used to describe the conditions under which whole-body plethysmograph Penh measurements can be used to detect changes in sRaw.
Subject(s)
Airway Resistance , Plethysmography, Whole Body , Pulmonary Ventilation , Respiration Disorders/diagnosis , Airway Resistance/drug effects , Algorithms , Animals , Bronchoconstrictor Agents/toxicity , Methacholine Chloride/toxicity , Models, Biological , Respiration Disorders/chemically induced , Respiration Disorders/physiopathology , Respiratory System/drug effects , Respiratory System/physiopathology , Severity of Illness IndexABSTRACT
This study compares basic respiratory variables (rate, tidal and minute volumes) with time-, flow- and ratio-derived parameters obtained using head-out plethysmography in rats following administration of reference drugs (isotonic saline, 2.0 mL/kg, IV; albuterol, 400 µg/kg, inhalation; methacholine, 136 µg/kg, IV; and remifentanil, 14 µg/kg, IV) to identify respiratory variables with superior sensitivity. Paired t-tests by block-period, and analysis of covariance (ANCOVA) with baseline as covariate and a posteriori pair-wise comparisons using Dunnett's test were used. Variations in respiratory parameters observed over time justify the use of a control group in any respiratory safety pharmacology study for inter-groups comparison. Handling-, and slumbering-, induced perturbations were minimal. The system was sensitive and specific to detect changes in respiratory variables related to pharmacologically-induced bronchodilation, bronchoconstriction and central respiratory depression. The standard variables (respiratory rate, tidal and minute volumes) confirmed to be the cornerstone of respiratory safety pharmacology to detect pharmacological changes. Flow-derived parameters appeared as highly valuable complement for interpretation of respiratory response, whereas time- and ratio-derived parameters presented limited added value during interpretation.
Subject(s)
Respiration/drug effects , Respiratory System/drug effects , Albuterol/administration & dosage , Albuterol/pharmacology , Albuterol/toxicity , Animals , Bronchoconstriction/drug effects , Consciousness , Dose-Response Relationship, Drug , Inspiratory Capacity/drug effects , Male , Methacholine Chloride/administration & dosage , Methacholine Chloride/pharmacology , Methacholine Chloride/toxicity , Piperidines/administration & dosage , Piperidines/pharmacology , Piperidines/toxicity , Plethysmography , Rats , Rats, Sprague-Dawley , Remifentanil , Reproducibility of Results , Respiratory Function Tests , Respiratory Insufficiency/chemically induced , Respiratory Rate/drug effects , Sodium Chloride/administration & dosage , Sodium Chloride/pharmacology , Sodium Chloride/toxicity , Tidal Volume/drug effectsABSTRACT
Transforming growth factor (TGF)-alpha and its receptor, the epidermal growth factor receptor, are induced after lung injury and are associated with remodeling in chronic pulmonary diseases, such as pulmonary fibrosis and asthma. Expression of TGF-alpha in the lungs of adult mice causes fibrosis, pleural thickening, and pulmonary hypertension, in addition to increased expression of a transcription factor, early growth response-1 (Egr-1). Egr-1 was increased in airway smooth muscle (ASM) and the vascular adventitia in the lungs of mice conditionally expressing TGF-alpha in airway epithelium (Clara cell secretory protein-rtTA(+/-)/[tetO](7)-TGF-alpha(+/-)). The goal of this study was to determine the role of Egr-1 in TGF-alpha-induced lung disease. To accomplish this, TGF-alpha-transgenic mice were crossed to Egr-1 knockout (Egr-1(ko/ko)) mice. The lack of Egr-1 markedly increased the severity of TGF-alpha-induced pulmonary disease, dramatically enhancing airway muscularization, increasing pulmonary fibrosis, and causing greater airway hyperresponsiveness to methacholine. Smooth muscle hyperplasia, not hypertrophy, caused the ASM thickening in the absence of Egr-1. No detectable increases in pulmonary inflammation were found. In addition to the airway remodeling disease, vascular remodeling and pulmonary hypertension were also more severe in Egr-1(ko/ko) mice. Thus, Egr-1 acts to suppress epidermal growth factor receptor-mediated airway and vascular muscularization, fibrosis, and airway hyperresponsiveness in the absence of inflammation. This provides a unique model to study the processes causing pulmonary fibrosis and ASM thickening without the complicating effects of inflammation.
Subject(s)
Bronchial Hyperreactivity/physiopathology , Early Growth Response Protein 1/physiology , Lung/pathology , Pulmonary Fibrosis/pathology , Transforming Growth Factor alpha/physiology , Airway Resistance , Albuterol/pharmacology , Animals , Bronchial Hyperreactivity/chemically induced , Bronchial Hyperreactivity/genetics , Cells, Cultured/drug effects , Cells, Cultured/pathology , Disease Models, Animal , Early Growth Response Protein 1/biosynthesis , Early Growth Response Protein 1/genetics , ErbB Receptors/antagonists & inhibitors , Fibroblasts/cytology , Humans , Hyperplasia , Lung Compliance , Methacholine Chloride/toxicity , Mice , Mice, Knockout , Mice, Transgenic , Muscle, Smooth/pathology , Muscle, Smooth, Vascular/drug effects , Muscle, Smooth, Vascular/pathology , Pulmonary Artery/cytology , Pulmonary Fibrosis/genetics , Pulmonary Fibrosis/physiopathology , Recombinant Fusion Proteins/physiology , Transforming Growth Factor alpha/adverse effects , Weight LossABSTRACT
Previously, we found pulmonary gas trapping to be a rapid, simple and objective measure of methacholine-induced airway obstruction in naïve mice. In this study we extended that finding by using methacholine-induced pulmonary gas trapping to differentiate airway responses of ovalbumin-sensitized, ovalbumin-exposed (Positive Control) and ovalbumin-sensitized, sodium chloride-exposed (Negative Control) mice. Additionally, pulmonary gas trapping and enhanced pause were compared following methacholine exposure in sensitized and nonsensitized mice. Finally, we examined by nose-only inhalation the ability of the glucocorticosteroid budesonide and the peroxisome proliferator-activated receptor-gamma agonist ciglitazone to modify methacholine-induced airway responses in ovalbumin-sensitized mice. Positive Controls exhibited a 7.8-fold increase in sensitivity and a 2.4-fold enhancement in the maximal airway obstruction to methacholine versus Negative Controls. Following methacholine, individual Positive and Negative Control mouse enhanced pause values overlapped in 9 of 9 studies, whereas individual Positive and Negative Control mouse excised lung gas volume values overlapped in only 1 of 9 studies, and log[excised lung gas volume] correlated (P=0.023) with in vivo log[enhanced pause] in nonsensitized mice. Finally, budesonide (100.0 or 1000.0 microg/kg) reduced methacholine-mediated airway responses and eosinophils and neutrophils, whereas ciglitazone (1000.0 microg/kg) had no effect on methacholine-induced pulmonary gas trapping, but reduced eosinophils. In conclusion, pulmonary gas trapping is a more reproducible measure of methacholine-mediated airway responses in ovalbumin-sensitized mice than enhanced pause. Also, excised lung gas volume changes can be used to monitor drug interventions like budesonide. Finally, this study highlights the importance of running a positive comparator when examining novel treatments like ciglitazone.
Subject(s)
Asthma/drug therapy , Bronchodilator Agents/pharmacology , Budesonide/pharmacology , Thiazolidinediones/pharmacology , Administration, Inhalation , Airway Obstruction/chemically induced , Airway Obstruction/drug therapy , Animals , Anti-Asthmatic Agents/administration & dosage , Anti-Asthmatic Agents/pharmacology , Bronchial Hyperreactivity/chemically induced , Bronchial Hyperreactivity/drug therapy , Bronchodilator Agents/administration & dosage , Budesonide/administration & dosage , Disease Models, Animal , Dose-Response Relationship, Drug , Eosinophils/drug effects , Eosinophils/metabolism , Male , Methacholine Chloride/toxicity , Mice , Mice, Inbred BALB C , Neutrophils/drug effects , Neutrophils/metabolism , Ovalbumin , PPAR gamma/agonists , Thiazolidinediones/administration & dosageABSTRACT
OBJECTIVES: This study compared the protective effect of two respiratory protection devices during exposure in a pig confinement building. METHODS: Thirty-six healthy persons were exposed for 3 hours in the building, 12 without any protection, 12 with a particle-filter mask, and 12 with a mask filtering both particles and gases. Symptoms, body temperature, nasal lavage fluid, exhaled nitric oxide, and bronchial responsiveness to methacholine were assessed before and after the exposure. Pre- and postexposure urine and blood samples were collected. RESULTS: After the exposure, the participants with respirators reported fewer symptoms than those without. Wearing a mask also reduced the inflammatory response assessed with nasal lavage (cell concentration, interleukins 6 and 8) and peripheral blood (cell number). Lung function was significantly impaired only in the unprotected group; postexposure vital capacity and forced expiratory volume in 1 second showed a decrease of 3-4% from the preexposure levels (P=0.006 and P=0.002, respectively). Bronchial responsiveness (P<0.01) and body temperature (P<0.001) increased similarly in the three groups. Bronchial responsiveness to methacholine increased 2.7, 2.4, and 2.1 doubling concentration steps for those unprotected, those using a particle-filter mask, and those using a mask with particle and gas filters, respectively. The prostaglandin D2 metabolite, 9a, 11b-PGF2 increased significantly (P=0.003) only in those unprotected. CONCLUSIONS: Wearing a respirator in a pig confinement building reduces the inflammatory reaction but does not influence the increase in bronchial responsiveness, with no difference between the use of a particle-filter mask or a mask with a particle-gas filter combination.
Subject(s)
Air Pollution, Indoor/adverse effects , Filtration/instrumentation , Gases , Inhalation Exposure/prevention & control , Occupational Exposure/prevention & control , Respiratory Protective Devices , Adult , Animals , Cytokines , Female , Humans , Male , Methacholine Chloride/toxicity , Middle Aged , Nitric Oxide/toxicity , Pneumonia/physiopathology , Pneumonia/prevention & control , Respiration Disorders/physiopathology , Respiration Disorders/prevention & control , Swine , Time FactorsABSTRACT
BACKGROUND: Although indoleamine 2,3-dioxygenase (IDO)-mediated immune suppression of mesenchymal stem cells (MSCs) has been revealed in septic and tumor microenvironments, the role of IDO in suppressing allergic airway inflammation by MSCs is not well documented. We evaluated the effects of adipose-derived stem cells (ASCs) on allergic inflammation in IDO-knockout (KO) asthmatic mice or asthmatic mice treated with ASCs derived from IDO-KO mice. METHODS AND FINDINGS: ASCs were injected intravenously in wild-type (WT) and IDO-KO asthmatic mice. Furthermore, asthmatic mice were injected with ASCs derived from IDO-KO mice. We investigated the immunomodulatory effects of ASCs between WT and IDO-KO mice or IDO-KO ASCs in asthmatic mice. In asthmatic mice, ASCs significantly reduced airway hyperresponsiveness, the number of total inflammatory cells and eosinophils in bronchoalveolar lavage fluid (BALF), eosinophilic inflammation, goblet hyperplasia, and serum concentrations of total and allergen-specific IgE and IgG1. ASCs significantly inhibited Th2 cytokines, such as interleukin (IL)-4, IL-5, and IL-13, and enhanced Th1 cytokine (interferon-γ) and regulatory cytokines (IL-10, TGF-ß) in BALF and lung draining lymph nodes (LLNs). ASCs led to significant increases in regulatory T-cells (Tregs) and IL-10+ T cell populations in LLNs. However, the immunosuppressive effects of ASCs did not significantly differ between WT and IDO-KO mice. Moreover, ASCs derived from IDO-KO mice showed immunosuppressive effects in allergic airway inflammation. CONCLUSIONS: IDO did not play a pivotal role in the suppression of allergic airway inflammation through ASCs, suggesting that it is not the major regulator responsible for suppressing allergic airway inflammation.
Subject(s)
Adipose Tissue/cytology , Asthma/genetics , Asthma/pathology , Indoleamine-Pyrrole 2,3,-Dioxygenase/genetics , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/immunology , Adipose Tissue/immunology , Animals , Asthma/therapy , Bronchoalveolar Lavage Fluid/cytology , Cell- and Tissue-Based Therapy/methods , Cells, Cultured , Eosinophils/cytology , Female , Goblet Cells/pathology , Hyperplasia/pathology , Immunoglobulin E/blood , Immunoglobulin G/blood , Inflammation/therapy , Interferon-gamma/metabolism , Interleukin-10/metabolism , Interleukin-13/metabolism , Interleukin-4/metabolism , Interleukin-5/metabolism , Lymphocyte Count , Methacholine Chloride/toxicity , Mice , Mice, Inbred C57BL , Mice, Knockout , T-Lymphocytes, Regulatory/cytology , T-Lymphocytes, Regulatory/immunology , Transforming Growth Factor beta/metabolismABSTRACT
Inhalation of high concentrations of sulfur dioxide (SO2) affects the lungs and can be immediately dangerous to life. We examined the development of acute and long-term effects after exposure of SO2 in Sprague-Dawley rats, in particular inflammatory responses, airway hyperresponsiveness (AHR) and lung fibrosis. Animals were subjected to a single exposure of 2200ppm SO2 during 10min and treated with a single dose of the anti-inflammatory corticosteroid dexamethasone 1h following exposure. Exposed rats showed labored breathing, decreased body-weight and an acute inflammation with neutrophil and macrophage airway infiltrates 5h post exposure. The acute effects were characterized by bronchial damage restricted to the larger bronchi with widespread injured mucosal epithelial lining. Rats displayed hyperreactive airways 24h after exposure as indicated by increased methacholine-induced respiratory resistance. The inflammatory infiltrates remained in lung tissue for at least 14 days but at the late time-point the dominating granulocyte types had changed from neutrophils to eosinophils. Analysis of immunoregulatory and pro-inflammatory cytokines in serum and airways implicated mixed macrophage phenotypes (M1/M2) and T helper cell activation of both TH1 and TH2 subtypes. Increased expression of the pro-fibrotic cytokine TGFß1 was detected in airways 24h post exposure and remained increased at the late time-points (14 and 28 days). The histopathology analysis confirmed a significant collagen deposition 14 days post exposure. Treatment with dexamethasone significantly counteracted the acute inflammatory response but was insufficient for complete protection against SO2-induced adverse effects, i.e. treatment only provided partial protection against AHR and the long-term fibrosis.
Subject(s)
Inflammation/drug therapy , Lung Injury/drug therapy , Pulmonary Fibrosis/drug therapy , Sulfur Dioxide/toxicity , Administration, Inhalation , Animals , Anti-Inflammatory Agents/pharmacology , Bronchial Hyperreactivity/chemically induced , Bronchial Hyperreactivity/drug therapy , Dexamethasone/pharmacology , Disease Models, Animal , Dose-Response Relationship, Drug , Eosinophils/drug effects , Eosinophils/metabolism , Female , Inflammation/chemically induced , Lung/drug effects , Lung/pathology , Lung Injury/chemically induced , Macrophages/drug effects , Macrophages/metabolism , Methacholine Chloride/toxicity , Neutrophils/drug effects , Neutrophils/metabolism , Pulmonary Fibrosis/chemically induced , Rats , Rats, Sprague-Dawley , Respiratory Hypersensitivity/chemically induced , Respiratory Hypersensitivity/drug therapy , Sulfur Dioxide/administration & dosage , T-Lymphocytes, Helper-Inducer/drug effects , T-Lymphocytes, Helper-Inducer/metabolism , Transforming Growth Factor beta1/genetics , Transforming Growth Factor beta1/metabolismABSTRACT
Clinical and experimental studies have reported that short-term exposure to particulate air pollution is associated with inflammation, oxidative stress and impairment of lung function. Emodin (1,3,8-trihydroxy-6-methylanthraquinone) has a strong antioxidant and anti-inflammatory actions. Therefore, in the present study, we evaluated the possible ameliorative effect of emodin on diesel exhaust particles (DEP)-induced impairment of lung function, inflammation and oxidative stress in mice. Mice were intratracheally instilled with DEP (20 µg/mouse) or saline (control). Emodin was administered intraperitoneally 1h before and 7h after pulmonary exposure to DEP. Twenty-four hours following DEP exposure, we evaluated airway resistance measured by forced oscillation technique, lung inflammation and oxidative stress. Emodin treatment abated the DEP-induced increase in airway resistance, and prevented the influx of neutrophils in bronchoalveolar lavage fluid. Similarly, lung histopathology confirmed the protective effect of emodin on DEP-induced lung inflammation. DEP induced a significant increase of proinflammatory cytokines in the lung including tumor necrosis factor α, interleukin 6 and interleukin 1ß. The latter effect was significantly ameliorated by emodin. DEP caused a significant increase in lung lipid peroxidation, reactive oxygen species and a significant decrease of reduced glutathione concentration. These effects were significantly mitigated by emodin. We conclude that emodin significantly mitigated DEP-induced increase of airway resistance, lung inflammation and oxidative stress. Pending further pharmacological and toxicological studies, emodin may be considered a potentially useful pulmonary protective agent against particulate air pollution-induced lung toxicity.
Subject(s)
Airway Resistance/drug effects , Emodin/pharmacology , Emodin/therapeutic use , Lung Diseases/drug therapy , Oxidative Stress/drug effects , Pneumonia/drug therapy , Protein Kinase Inhibitors/therapeutic use , Animals , Cytokines/metabolism , Disease Models, Animal , Dose-Response Relationship, Drug , Glutathione/metabolism , Lipid Peroxidation/drug effects , Lung/metabolism , Lung Diseases/chemically induced , Lung Diseases/pathology , Methacholine Chloride/toxicity , Mice , Mice, Inbred BALB C , Muscarinic Agonists/toxicity , Particulate Matter/toxicity , Pneumonia/chemically induced , Protein Kinase Inhibitors/pharmacology , Reactive Oxygen Species/metabolism , Time FactorsABSTRACT
The aim of this study was to compare the antianginal effects of two compounds that release nitric oxide (NO) spontaneously, i.e. (+/-)-N-[(E)-4-ethyl-3-[(Z-hydroxyimino]-5-nitro-3-hexenyl] -3-pyridinecarboxamide (FR144420) and (+/-)-(E)-ethyl-2-[(E)-hydroxyimino] -5-nitro-3-hexenamide (FK409), in two different rat models of coronary vasospasm. In the rat methacholine-induced coronary vasospasm model, FR144420 suppressed the elevation of the ST segment dose dependently and significantly at 1.0 mg/kg, i.d. 185 min after its administration. FK409 suppressed the ST elevation only 5 min after its administration at 1.0 mg/kg, i.d. FR144420 and FK409 significantly decreased mean blood pressure at all doses tested only 5 min after their intraduodenal administration, but did not change heart rate at any time. Although the suppression of the ST elevation by FK409 had the same duration as its hypotensive effect, the FR144420-induced suppression of the ST elevation lasted longer than its hypotensive effect. In the rat vasopressin-induced coronary vasospasm model, FR144420 (32 mg/kg) significantly inhibited the depression of the ST segment both 60 min and 120 min after oral administration, whereas FK409 (32 mg/kg) significantly inhibited this ST depression only 60 min after oral administration. These data suggest that FR144420 inhibits coronary vasospasm for longer than FK409 does and particularly shows more prolonged antianginal effects than hypotensive effects in the methacholine-induced coronary vasospasm model. Thus FR144420 is expected to be a useful NO releaser for investigating the in vivo actions of NO.
Subject(s)
Coronary Vasospasm/drug therapy , Nicotinic Acids/therapeutic use , Nitro Compounds/therapeutic use , Vasodilator Agents/therapeutic use , Animals , Blood Pressure/drug effects , Coronary Vasospasm/chemically induced , Electrocardiography/drug effects , Male , Methacholine Chloride/toxicity , Nitric Oxide/chemistry , Rats , Rats, Sprague-Dawley , Vasopressins/toxicityABSTRACT
Trimellitic anhydride (TMA) is a low-molecular-weight chemical known to cause occupational asthma. The dose-response study was designed to determine whether respiratory responses during a single inhalation challenge with TMA (25-30 mg/m3 for 30 min, 3 weeks after the initial induction), the ensuing non-specific airway hyperresponsiveness (AH) to methacholine (MCh) aerosol, and infiltration of eosinophilic granulocytes into the lungs of sensitized Brown Norway (BN) rats are associated and dependent on the concentration of TMA used for topical induction. The initial topical exposure concentrations were 1, 5, and 25% TMA in acetone:olive oil (AOO) followed by a booster induction 1 week later. In the time course study BN rats received AOO alone or were sensitized to the minimal sensitizing topical concentration of TMA (5%) and were the subsequently challenged with TMA on Days 17, 24, 41, 47, 55, and 66, followed by a MCh challenge 1 day later. One additional group of rats was sensitized to 5% TMA but were repeatedly challenged with MCh without prior TMA challenge. In the dose-response study the rats sensitized topically to TMA (5 and 25% in AOO) displayed unequivocal changes in breathing patterns upon challenge with TMA, including an increased responsiveness to MCh aerosol. These findings were associated with a sustained pulmonary eosinophilic inflammation. All endpoints demonstrated consistently that 5% TMA in AOO constitutes the minimal sensitizing concentration. When rats were topically sensitized with this concentration and repeatedly challenged with TMA over a time period of 7 weeks, it became apparent that challenge exposures in BN rats may be false negative when performed at time periods less than 3 weeks after the initial induction. Despite the time-related increased responsiveness elicited by the repeated TMA challenge exposures, the MCh challenge revealed increased non-specific airway hyperreactivity exclusively on Day 17. After the sixth TMA-challenge, the respiratory response and lung weights of rats sensitized topically were essentially similar to those observed in the repetitively re-challenged control group (induction: vehicle only; repeated booster challenge exposures with TMA). Thus, it appears, that in this animal model the effective concentration for successful topical sensitization must be at least approximately 5%. The repeated low-dose re-challenge with TMA in topically sensitized rats resulted in similar or slightly aggravated time-related responses over a period of 7 weeks. An over-proportionally increased susceptibility of rats receiving a topical priming dose prior to repeated inhalation challenge exposures was not observed. In summary, this study shows that the analysis of functional changes in breathing patterns is suitable to identify respiratory allergy. Repeated short-term inhalation exposures to mildly irritant concentrations (but low doses) of chemical asthmagens may be of higher concern than topical exposures.
Subject(s)
Allergens/toxicity , Phthalic Anhydrides/toxicity , Respiratory Hypersensitivity/chemically induced , Administration, Topical , Aerosols , Allergens/immunology , Animals , Bronchoconstrictor Agents/immunology , Bronchoconstrictor Agents/toxicity , Dose-Response Relationship, Immunologic , Lung/drug effects , Lung/immunology , Lung/pathology , Lymph Nodes/drug effects , Lymph Nodes/immunology , Lymph Nodes/pathology , Male , Methacholine Chloride/immunology , Methacholine Chloride/toxicity , Organ Size/drug effects , Phthalic Anhydrides/immunology , Rats , Rats, Inbred BN , Respiratory Hypersensitivity/immunology , Respiratory Hypersensitivity/pathology , Respiratory Mechanics/drug effects , Respiratory Mechanics/immunology , Time FactorsABSTRACT
Removal of the epithelial layer of rat tracheal tissue did not affect the methacholine-induced contraction of the tracheal smooth muscle, but attenuated the (-)-isoprenaline induced relaxation (expressed as percentage of the methacholine contraction). In this way the epithelial layer seemed to play a role in the maintenance of an autonomic balance between sympathetic and parasympathetic receptor responses. Incubation of rat tracheal tissue with cumene hydroperoxide (3 x 10(-5)-10(-3) M) resulted in a dose-dependent destruction and (partial) removal of the epithelial layer. Cumene hydroperoxide diminished muscarinic receptor responses of the rat trachea. Moreover, the autonomic balance between muscarinic and beta-adrenoceptor responses was affected. The effects of cumene hydroperoxide on receptor responses were more pronounced after epithelium removal. The protective role of the epithelial layer of pulmonary tissue against oxidative stress has therefore been emphasized.
Subject(s)
Benzene Derivatives/toxicity , Isoproterenol/toxicity , Methacholine Chloride/toxicity , Muscle, Smooth/drug effects , Receptors, Adrenergic, beta/drug effects , Receptors, Muscarinic/drug effects , Trachea/drug effects , Animals , Epithelium/physiology , Male , Muscle Contraction/drug effects , Rats , Rats, Inbred Strains , Trachea/pathologyABSTRACT
The effects of histamine and methacholine aerosols and of a fixed inspiratory resistance on tidal breathing flow-volume loops (TBFVL) were investigated using 18 unsedated, standing, healthy thoroughbred horses. The data were first analysed using traditional flow-volume loop indices and then reduced using standardized factor scoring coefficients obtained in a previous study in this laboratory using similar experimental techniques. On the basis of resting TBFVL analysis, the degree of pulmonary dysfunction caused by inhalation of histamine and methacholine aerosols with concentrations of 10 and 2 mg/ml, respectively, was similar. The fixed resistance also caused significant changes in the resting spirogram and TBFVL indices, suggesting that this model may prove valuable for further studies involving upper respiratory tract (URT) conditions. Administration of histamine and methacholine aerosols resulted in significant changes in all factor scores, although most of the observed changes were due to the effects of these aerosols on the respiratory rate. These findings re-emphasize the importance of the effects of respiratory rate on pulmonary mechanics. Application of the resistance resulted in significant changes in factor score 3, the 'inspiratory' factor, which lends support to the validity of this model for URT conditions. The close agreement between the factor scores obtained under controlled conditions in this study and in a previous study in this laboratory confirms that the factor analysis used for both of these studies provides an adequate means of reducing TBFVL data obtained from thoroughbred horses. The large intra- and inter-individual variation observed both with the indices of TBFVL and with the factor scores limits the potential of these variables for detecting individual animals with obstructive airway disease. Re-evaluation of these indices under the stress of exercise may reduce the variability observed in these data and may increase the magnitude of differences between different animals, providing a means of detecting individual animals with subclinical obstructive airway conditions.