Bronchoalveolar lavage analysis

Bronchoalveolar lavages (BAL) of rat lungs were performed in five animals of each dose group at all five time points. The method is based on Henderson et al. [57] with minor modifications. Lungs were lavaged twice using 4 mL 0.9% NaCl. The following parameters were determined from collected lavage fluids: total cell count, differential cell count (macrophages, neutrophils, eosinophils and lymphocytes), biochemical mediators (lactic dehydrogenase, ß-glucuronidase and total protein), as well as cytokine levels. Total cell counts were measured using a counting chamber (Fuchs-Rosenthal). Differential cell counts were prepared by centrifugation of BAL fluid on cytoslides and subsequent Giemsa staining. Biochemical indicators were determined in the supernatant of centrifuged BAL fluid according to routine clinical chemistry protocols.