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Equine precision cut lung slices (PCLS) were used to evaluate the ability of several pharmacologic compounds, parasympatholytics atropine, ipratropium, tiotropium and phosphodiesterase (PDE) 4-inhibitor cilomilast, to inhibit methacholine (MCh) induced bronchial constriction. The study war performed using vital lung samples from 20 horses, harvested immediately after euthanasia and lung examination.Tissue samples from the cranial, caudal and accessory lobe were taken for histological examination. Viability in PCLS of 250 +/- 20 microm was confirmed by observation of cilia movement and bronchial contraction in response to the addition of increasing concentrations of MCh (10(-8) to 10(-4) mol/l). Washing in pure RPMI-medium resulted in bronchial reopening. After incubation (20 minutes in an RPMI-solution containing cilomilast (10(-5) mol/l), atropine, ipratropium or tiotropium (10(-7) mol/l) each group (randomized, placebo controlled) was treated with MCh again. After reopening the unaffected viability of the PCLS was confirmed by the addition of MCh. EC50 values were calculated from bronchial cross-sections for each contraction. EC50 values significantly increased after atropine, ipratropium and tiotropium addition. No difference was found for cilomilast and in control slices. Statistical analysis showed a large interindividual, but a small intraindivual variability. No correlation was found between the clinical examination, histological findings and PCLS' responsiveness. We were able to confirm Vietmeier's (2007) values for the responsiveness of equine PCLS to MCh and to proof the effect of several new bronchodilatators using a new technique that allows the assessment of several substances in the same horse.