Implantação de técnicas histológicas no laboratório de imunofarmacologia /UFPB: aspectos morfológicos de desordens respiratórias experimentais

Abstract

The histological technique is a set of procedures based on the preparation of samples for analysis under microscopy of normal or pathological elements. Pre clinical work developed at the immunopharmacology laboratory of the UFPB, demonstrated the need for implantation of the histological technique for morphological studies in models of respiratory disorders. Respiratory diseases such as allergic diseases and acute lung injury are seen as a global health problem resulting from a complex interaction between genetic and environmental factors. Allergic diseases are characterized by increased mucus production, eosinophilic inflammation, bronchial hyperresponsiveness and if more severe tissue remodeling. On the other hand acute lung injury is affected by involvement of the alveolar region involving epithelial and endothelial cell, influx of inflammatory cells, discontinuation of surfactant production and pulmonary edema. Histological staining is useful in observing these parameters for example Hematoxylin eosin (H & E) demonstrates non specific parameters such as infiltration and edema, Schiff's Periodic Acid (PAS) reveals goblet cells producing mucus, Toluidine Blue (AT) emphasizes metachromatic mast cells and the Gomori Tricomo (TG) shows the collagen fibers. The purpose of this work was to implant the histological techniques to evaluate and differentiate in morphological terms the respiratory disorder models developed in the immunopharmacology laboratory. In the model of allergic diseases of the airways, physiological BALB / c mice (Basal), sensitized and challenged with OVA (OVA) or treated with budesonide (BUD) were used. In the model of Acute Lung Injury (LPA) the mice were challenged with LPS (LPS) or treated with dexamethasone (DEXA). On the last day of the whole protocol of animals with euthanasia and were collected the nasal fluid lavage (NALF), bronchoalveolar lavage (BAL), lung and nasal cavity. A statistical analysis was done by (ANOVA) one-way using GraphPad Prism® version 5.01. In the model of airway allergic diseases, the OVA group shows a significant increase (p <0.001) in the amount of NALF cells (0, 27 ± 0.02) and BAL cells (8.20 ± 0.08) when compared to the basal group (0.08 ± 0.01) (1.00 ± 0.15). The infiltrate was of the eosinophilic type (0.15 ± 0.015) (3.18 ± 0.53). A histological analysis of the nasal cavity of the OVA group showed H & E staining, cell infiltration (3.40 ± 0.31), by PAS, hypertrophy and hyperplasia of mucus producing cells (3.60 ± 0.24), by AT, increased mast cells (3.20 ± 0.37) and TG, subtle tissue remodeling (2.40 ± 0.40). In lung tissue, were increased cellular infiltration (3.80 ± 0.20) and the amount of mucus (3.60 ± 0.24) in the OVA group increased. Budesonide decreased these parameters. On LPA model, the LPS group showed increase of total cells (7.30 ± 0.26) and neutrophils (5.48 ± 0.41) compared to the basal group (0.32 ± 0.10) (0, 09 ± 0.02). A histological analysis showed H & E interstitial inflammation (3.60 ± 0.24), pulmonary edema (3.20 ± 0.37), hemorrhage (3.00 ± 0.31) and by TG, Collagen fibers (2.40 ± 0.24) in the LPS group compared to the basal group. With the implementation of histological techniques, it was possible to observe the participation of tissue components such as cells, extracellular matrix and mediators in respiratory disease models contributing to the pré clinical work developed in the immunopharmacology laboratory.