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Vol. 45. Issue 9.
Pages 422-428 (September 2009)
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Vol. 45. Issue 9.
Pages 422-428 (September 2009)
Original Article
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Development of a Murine Model of Airway Inflammation and Remodeling in Experimental Asthma
Desarrollo de un modelo murino de inflamación y remodelación de vías respiratorias en asma experimental
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Rebeca Fraga-Iriso, Laura Núñez-Naveira, Nadia S. Brienza, Alberto Centeno-Cortés, Eduardo López-Peláez, Héctor Verea, David Ramos-Barbón
Corresponding author
David.Ramos.Barbon@sergas.es

Corresponding author.
Unidad de Investigación Respiratoria, Servicio de Neumología, Complejo Hospitalario Universitario, Instituto de Investigación Biomédica de A Coruña (INIBIC), A Coruña, Spain
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Article information
Abstract
Background and Objective

Experimental animal models are necessary for studying asthma disease mechanisms and for identifying new therapeutic targets. We present a murine model of experimental asthma that allows integrated, quantitative assessment of airway inflammation and remodeling.

Material and Methods

BALB/c mice were sensitized to ovalbumin (OVA) and challenged with OVA or vehicle 3 times per week for 12 weeks.

Results

On bronchoalveolar lavage, the OVA-sensitized mice had significantly higher total leukocyte counts, with a median (Q25-Q75) of 670.0 cells/mL×10-3 (376.2-952.5) in comparison with 40.0 cells/ mL×10-3 (60.0-85.0) in controls (P=.001), and higher eosinophil and differential lymphocyte counts. In sagittal sections of lungs inflated to a standard pressure, the OVA-sensitized animals showed goblet cell hyperplasia in the respiratory epithelium (periodic acid-Schiff staining, 53.89 [36.26-62.84] cells/mm2 vs 0.66 [0.00-1.06] cells/mm2, P<.001), dense mononuclear and eosinophilic inflammatory infiltrates (hematoxylin-eosin, 32.87 [27.34-37.13] eosinophils/mm2 vs 0.06 [0.00-0.20] eosinophils/mm2, P=.002), subepithelial infiltration by mast cells (toluidine blue, 2.88 [2.00-3.28] mast cells/mm2 vs 0.28 [0.15-0.35] mast cells/mm2, P<.001), increased contractile tissue mass (immunofluorescence analysis for α-smooth-muscle actin, 2.60 [2.28-2.98] vs 1.08 [0.93-1.16], dimensionless, P<.001) and enhanced extracellular matrix deposition (Masson's trichrome, 2.18 [1.85-2.80] vs 0.50 [0.37-0.65], dimensionless, P<.001).

Conclusions

Our dataset describes an experimental model of asthma which is driven by prolonged allergen exposure and in which airway inflammation and remodeling develop and are assessed together.

Keywords:
Asthma
Animal disease models
Inflammation
Goblet cells
Eosinophils
Lymphocytes
Mast cells
Smooth muscle
Extracellular matrix
Resumen
Introducción

La investigación de los mecanismos de enfermedad del asma y la identificación de nuevas dianas terapéuticas requieren modelos animales experimentales. En este trabajo presentamos los datos del desarrollo de un modelo murino de asma experimental que permite valorar de forma conjunta parámetros de inflamación y remodelación de las vías respiratorias mediante morfología cuantitativa.

Material y métodos

Se sensibilizó a ovoalbúmina a ratones Balb/c y se les realizó broncoprovocación con ovoalbúmina o excipiente 3 veces por semana durante 12 semanas.

Resultados

En el lavado broncoalveolar, los ratones del grupo de ovoalbúmina presentaron un incremento significativo de leucocitos totales, con una mediana (cuartiles 25–75) de 670,0 células/ml·103 (376,2–952,5), frente a 40,0 células/ml·103 (60,0–85,0) en controles (p = 0,001), y de las fracciones eosinófila y linfocitaria en recuento diferencial. En secciones sagitales de los pulmones inflados a presión estandarizada, estos ratones mostraron hiperplasia de células caliciformes en el epitelio respiratorio —reacción de ácido peryódico de Schiff: 53,89 (36,26—62,84) frente a 0,66 (0,00–1,06) células/mm2 (p < 0,001)—, densa infiltración inflamatoria mononuclear y eosinófila —hematoxilina-eosina: 32,87 (27,34–37,13) frente a 0,06 (0,00–0,20) eosinófilos/mm2 (p = 0,002)—, infiltración subepitelial por mastocitos —azul de toluidina: 2,88 (2,00–3,28) frente a 0,28 (0,15–0,35) mastocitos/mm2 (p < 0,001)—, incremento de la masa de tejido contráctil —inmunofluorescencia para alfaactina de músculo liso: 2,60 (2,28–2,98) frente a 1,08 (0,93–1,16), adimensional (p<0,001)— e incremento del depósito de matriz extracelular (tricrómico de Masson: 2,18 (1,85–2,80) frente a 0,50 (0,37–0,65), adimensional (p < 0,001)—.

Conclusiones

Los datos aportados configuran un modelo de asma experimental inducida por exposición alergénica prolongada, con desarrollo y evaluación integrada de inflamación y remodelación de vías respiratorias.

Palabras clave:
Asma
Modelos animales de enfermedad
Inflamación
Células caliciformes
Eosinófilos
Linfocitos
Mastocitos
Músculo liso
Matriz extracelular
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Copyright © 2009. Sociedad Española de Neumología y Cirugía Torácica
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