Effects of Roflumilast, Selective PDE4 Inhibitor, on Airway Reactivity in Ovalbumin-Sensitized Guinea Pigs

Abstract Background: Roflumilast as a phosphodiesterase 4 inhibitor has shown to increase lung functions and decrease the number of exacerbations in chronic obstructive lung disease. In this study, its ability to decrease the airway hyperresponsiveness in a model of eosinophil inflammation was evaluated. Methods: Healthy adult male guinea pigs were divided into groups as follows: the first group was considered as a healthy control group (without sensitization and therapy), animals in the second group were sensitized with ovalbumin, but left without further treatment, and the animals in the third group were sensitized with ovalbumin and treated with roflumilast perorally for 7 consecutive days. In vivo airway reactivity was evaluated using double-chamber whole body plethysmograph and measuring the specific airway resistance after nebulization of histamine aerosol. In vitro experiments were performed with tissue strips of trachea and lungs in organ bath, where their contractile responses to cumulative doses of acetylcholine and histamine were registered. The numbers of inflammatory cells in blood and bronchoalveolar lavage fluid were measured using standard staining. Results: Guinea pigs with roflumilast treatment showed decreased in vivo and in vitro airway reactivity associated with suppressed recruitment of inflammatory cells (especially eosinophils) in blood and bronchoalveolar lavage fluid. Conclusion: Roflumilast has demonstrated the therapeutic potential in the model of ovalbumin induced eosinophil inflammation typically present in patients with bronchial asthma.

Download Full-text

Influence of sensitization and allergen provocation procedures on the development of allergen-induced bronchial hyperreactivity in conscious, unrestrained guinea-pigs

Mediators of Inflammation ◽

10.1155/s0962935195000263 ◽

1995 ◽

Vol 4 (2) ◽

pp. 149-156 ◽

Cited By ~ 2

Author(s):

R. E. Santing ◽

C. G. Olymulder ◽

B. van Diepen ◽

H. Meurs ◽

J. Zaagsma

Keyword(s):

Bronchoalveolar Lavage ◽

Guinea Pigs ◽

Bronchoalveolar Lavage Fluid ◽

Late Phase ◽

Inflammatory Cells ◽

Lavage Fluid ◽

Bronchial Hyperreactivity ◽

Cellular Activation ◽

Allergen Provocation ◽

Similar Development

The effects of different sensitization and allergen provocation regimens on the development of allergen-induced bronchial hyperreactivity (BHR) to histamine were investigated in conscious, unrestrained guinea-pigs. Similar early and late phase asthmatic reactions, BHR for inhaled histamine after the early (6 h) as well as after the late reaction (24 h), and airway inflammation were observed after a single allergen provocation in animals sensitized to produce mainly IgG or IgE antibodies, respectively. Repeating the allergen provocation in the IgE-sensitized animals after 7 days, using identical provocation conditions, resulted in a similar development of BHR to histamine inhalation. Repetition of the allergen provocation during 4 subsequent days resulted in a decreased development of BHR after each provocation, despite a significant increase in the allergen provocation dose necessary to obtain similar airway obstruction. The number of inflammatory cells in the bronchoalveolar lavage was not significantly changed after repeated provocation, when compared with a single allergen provocation. Finally, we investigated allergen-induced bronchial hyperreactivity by repetition of the sensitization procedure at day 7 and 14 (booster), followed by repeated allergen provocation twice a week for 5 weeks. Surprisingly, no BHR to histamine could be observed after either provocation, while the number of inflammatory cells in the bronchoalveolar lavage fluid after 5 weeks was enhanced compared with controls. These data indicate that both IgE and IgG sensitized guinea-pigs may develop bronchial hyperreactivity after a single allergen provocation. Repeated allergen exposure of IgE sensitized animals causes a gradual fading of the induced hyperreactivity despite the on-going presence of inflammatory cells in the airways, indicating a mechanism of reduced cellular activation.

Download Full-text

Cigarette smoking, emphysema, and damage to alpha 1-proteinase inhibitor

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1994.266.6.l593 ◽

1994 ◽

Vol 266 (6) ◽

pp. L593-L611 ◽

Cited By ~ 13

Author(s):

M. D. Evans ◽

W. A. Pryor

Keyword(s):

Cigarette Smoke ◽

Proteinase Inhibitor ◽

Inflammatory Cells ◽

Lavage Fluid ◽

Lung Lavage ◽

Tissue Destruction ◽

Phagocytic Cells ◽

Lung Fluid

The proteinase-antiproteinase theory for the pathogenesis of emphysema proposes that the connective tissue destruction associated with emphysema arises from excessive proteinase activity in the lower respiratory tract. For this reason, the relative activities of neutrophil elastase and alpha 1-proteinase inhibitor (alpha 1-PI) are considered important. Most emphysema is observed in smokers; therefore, alpha 1-PI has been studied as a target for smoke-induced damage. Damage to alpha 1-PI in lung fluid could occur by several mechanisms involving species delivered to the lung by cigarette smoke and/or stimulated inflammatory cells. Oxidative damage to alpha 1-PI has received particular attention, since both cigarette smoke and inflammatory cells are rich sources of oxidants. In this article we review almost two decades of research on mechanistic studies of damage to alpha 1-PI by cigarette smoke and phagocytic cells in vitro, studies emphasizing the importance of elastinolytic activity in the pathogenesis of emphysema in vivo and studies of human lung lavage fluid to detect defects in alpha 1-PI at the molecular and functional levels.

Download Full-text

Bronchoalveolar Lavage Fluid Utilized Ex Vivo to Validate In Vivo Findings: Inhibition of Gap Junction Activity in Lung Tumor Promotion is Toll-Like Receptor 4-Dependent

Journal of Molecular Biomarkers & Diagnosis ◽

10.4172/2155-9929.1000160 ◽

2013 ◽

Vol 05 (01) ◽

Cited By ~ 2

Author(s):

Ross S Osgood

Keyword(s):

Bronchoalveolar Lavage ◽

Gap Junction ◽

Bronchoalveolar Lavage Fluid ◽

Lung Tumor ◽

Ex Vivo ◽

Tumor Promotion ◽

Lavage Fluid ◽

Toll Like Receptor ◽

Toll Like Receptor 4

Download Full-text

PAMP is a novel inhibitor of the tachykinin release in the airway of guinea pigs

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1997.272.6.l1066 ◽

1997 ◽

Vol 272 (6) ◽

pp. L1066-L1069

Author(s):

H. Kanazawa ◽

H. Kamoi ◽

T. Kawaguchi ◽

S. Shoji ◽

T. Fujii ◽

...

Keyword(s):

Substance P ◽

Bronchoalveolar Lavage ◽

Guinea Pigs ◽

Bronchoalveolar Lavage Fluid ◽

Lavage Fluid ◽

Neurokinin A ◽

Dependent Manner ◽

C Fiber ◽

Dose Dependent

Proadrenomedullin NH2-terminal 20 peptide (PAMP), a newly identified hypotensive peptide, may play physiological roles in airway and cardiovascular controls. This study was designed to determine the mechanism responsible for the bronchoprotective effects of PAMP on capsaicin-induced bron-choconstriction in anesthetized guinea pigs. PAMP (10(-8)-10(-6) M) significantly inhibited capsaicin-induced bronchoconstriction in a dose-dependent manner. The bronchoprotective effect of PAMP (10(-6) M) was as large as that of isoproterenol (10(-7) M) and lasted > 10 min. The concentration of immunoreactive substance P (SP) in bronchoalveolar lavage fluid after administration of capsaicin (4 x 10(-6) M) was 120 +/- 10 fmol/ml. PAMP significantly inhibited the release of immunoreactive SP in a dose-dependent manner (60 +/- 6 fmol/ml for (10(-6) M PAMP, P < 0.01; 84 +/- 6 fmol/ml for 10(-7) M PAMP, P < 0.01; and 95 +/- 6 fmol/ml for 10(-8) M PAMP, P < 0.05). PAMP (10(-6) M) did not significantly affect exogenous neurokinin A (NKA) or NKA + SP-induced bronchoconstriction, whereas isoproterenol (10(-7) M) significantly inhibited exogenous tachykinin-induced bronchoconstriction. These findings suggest that the bronchoprotective effects of PAMP are mainly due to inhibition of the release of tachykinins at airway C-fiber endings.

Download Full-text

Concentration-Time Models for the Effects of Ozone on Bronchoalveolar Lavage Fluid Protein from Rats and Guinea Pigs

Inhalation Toxicology ◽

10.3109/08958379209145300 ◽

1992 ◽

Vol 4 (1) ◽

pp. 1-16 ◽

Cited By ~ 3

Author(s):

Jerry W. Highfill ◽

Gary E. Hatch ◽

Ralph Slade ◽

Kay M. Crissrnan ◽

Joel Norwood ◽

...

Keyword(s):

Bronchoalveolar Lavage ◽

Guinea Pigs ◽

Bronchoalveolar Lavage Fluid ◽

Lavage Fluid ◽

Concentration Time

Download Full-text

Anti-inflammatory effects of zinc and alterations in zinc transporter mRNA in mouse models of allergic inflammation

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00280.2006 ◽

2007 ◽

Vol 292 (2) ◽

pp. L577-L584 ◽

Cited By ~ 80

Author(s):

Carol Lang ◽

Chiara Murgia ◽

Mary Leong ◽

Lor-Wai Tan ◽

Giuditta Perozzi ◽

...

Keyword(s):

Bronchoalveolar Lavage ◽

Bronchoalveolar Lavage Fluid ◽

Gene Array ◽

Allergic Inflammation ◽

Inflammatory Cells ◽

Lavage Fluid ◽

Mrna Levels ◽

Solute Carrier Family ◽

Zn Uptake ◽

Solute Carrier

There is clinical evidence linking asthma with the trace element, zinc (Zn). Using a mouse model of allergic inflammation, we have previously shown that labile Zn decreases in inflamed airway epithelium (Truong-Tran AQ, Ruffin RE, Foster PS, Koskinen AM, Coyle P, Philcox JC, Rofe AM, Zalewski PD. Am J Respir Cell Mol Biol 27: 286–296, 2002). Moreover, mild nutritional Zn deficiency worsens lung function. Recently, a number of proteins belonging to the Solute Carrier Family 39 (ZIP) and Solute Carrier Family 30 (ZnT) have been identified that bind Zn and regulate Zn homeostasis. Mice were sensitized, and subsequently aerochallenged, with ovalbumin to induce acute and chronic airway inflammation. Mice received 0, 54, or 100 μg of Zn intraperitoneally. Tissues were analyzed for Zn content and histopathology. Inflammatory cells were counted in bronchoalveolar lavage fluid. Cytokine and Zn transporter mRNA levels were determined by cDNA gene array and/or real-time PCR. Zn supplementation decreased bronchoalveolar lavage fluid eosinophils by 40 and 80%, and lymphocytes by 55 and 66%, in the acute and chronic models, respectively. Alterations in Zn transporter expression were observed during acute inflammation, including increases in ZIP1 and ZIP14 and decreases in ZIP4 and ZnT4. Zn supplementation normalized ZIP1 and ZIP14, but it did not affect mRNA levels of cytokines or their receptors. Our results indicate that inflammation-induced alterations in Zn transporter gene expression are directed toward increasing Zn uptake. Increases in Zn uptake may be needed to counteract the local loss of Zn in the airway and to meet an increased demand for Zn-dependent proteins. The reduction of inflammatory cells by Zn in the airways provides support for Zn supplementation trials in human asthmatic individuals.

Download Full-text

OKY-046 inhibits anaphylactic bronchoconstriction and reduces histamine level in bronchoalveolar lavage fluid in sensitized guinea pigs

Prostaglandins ◽

10.1016/0090-6980(90)90023-o ◽

1990 ◽

Vol 39 (6) ◽

pp. 623-637 ◽

Cited By ~ 9

Author(s):

F. Nambu ◽

S. Murakata ◽

T. Shiraji ◽

N. Omawari ◽

M. Sawada ◽

...

Keyword(s):

Bronchoalveolar Lavage ◽

Guinea Pigs ◽

Bronchoalveolar Lavage Fluid ◽

Lavage Fluid ◽

Histamine Level

Download Full-text

Increased levels of endothelin-1 in bronchoalveolar lavage fluid from patients with systemic sclerosis contribute to fibroblast mitogenic activity in vitro.

American Journal of Respiratory Cell and Molecular Biology ◽

10.1165/ajrcmb.11.4.7917311 ◽

1994 ◽

Vol 11 (4) ◽

pp. 439-445 ◽

Cited By ~ 94

Author(s):

A D Cambrey ◽

N K Harrison ◽

K E Dawes ◽

A M Southcott ◽

C M Black ◽

...

Keyword(s):

Systemic Sclerosis ◽

Bronchoalveolar Lavage ◽

Bronchoalveolar Lavage Fluid ◽

Lavage Fluid ◽

Mitogenic Activity ◽

Endothelin 1

Download Full-text

The effect of induced sputum and bronchoalveolar lavage fluid from patients with chronic obstructive pulmonary disease on neutrophil migration in vitro

Medicina ◽

10.3390/medicina46050044 ◽

2010 ◽

Vol 46 (5) ◽

pp. 315 ◽

Cited By ~ 4

Author(s):

Agnė Babušytė ◽

Jolanta Jeroch ◽

Rimantas Stakauskas ◽

Kristina Stravinskaitė ◽

Kęstutis Malakauskas ◽

...

Keyword(s):

Bronchoalveolar Lavage ◽

Bronchoalveolar Lavage Fluid ◽

Neutrophil Migration ◽

Lavage Fluid ◽

Interleukin 8 ◽

Induced Sputum ◽

Chronic Obstructive ◽

Healthy Individuals ◽

Copd Patients

Objective. The aim of study was to investigate a chemotactic effect of induced sputum and bronchoalveolar lavage fluid on blood neutrophils in patients with chronic obstructive pulmonary disease (COPD) and healthy individuals. Material and methods. Forty-three smokers with COPD, 19 ex-smokers with COPD, 13 healthy smokers, and 17 healthy nonsmokers were recruited to the study. Neutrophils were isolated from peripheral blood of study individuals. For the same experimental conditions, pooled induced sputum and bronchoalveolar lavage fluid of 20 COPD patients were used. Neutrophil chemotaxis in vitro was performed in cell-transmigration chamber. Substances tested for chemoattraction (interleukin-8, induced sputum, bronchoalveolar lavage fluid directly or in addition to interleukin-8) were added to lower wells. Upper wells were filled with 2.5×106/mL of neutrophil culture and incubated for 2 hours. Migration was analyzed by flow cytometry. Results. Interleukin-8 (10–100 ng/mL) induced a dose-dependant neutrophil migration in all the groups. Only 100 ng/L of interleukin-8 induced more intensive chemotaxis of neutrophils from COPD smokers as compared to ex-smokers (P<0.05). Such difference between healthy individuals was obtained using 30 ng/mL of interleukin-8 (P<0.05). Induced sputum/interleukin-8 (10–100 ng/mL), as well as induced sputum directly, induced neutrophil migration (P<0.05). Chemotaxis of neutrophils isolated from COPD patients and healthy nonsmokers did not depend on additional interleukin-8 concentration. Bronchoalveolar lavage fluid/interleukin-8 (30–100 ng/mL) induced more intensive migration of neutrophils from COPD patients than bronchoalveolar lavage fluid (P<0.05) alone. Conclusions. Migration of neutrophils isolated from patients with COPD was more intensive compared to healthy individuals. Induced sputum and bronchoalveolar lavage fluid directly and with addition of interleukin-8 stimulated chemotaxis, and it was higher in neutrophils from COPD patients. Migration of neutrophils did not depend on smoking status.

Download Full-text