Barton Ann Kristin, Niedorf Frank, Gruber Achim D, Kietzmann Manfred, Ohnesorge Bernhard
Clinic for Horses, University of Veterinary Medicine Hanover, Foundation, Germany.
Berl Munch Tierarztl Wochenschr. 2010 May-Jun;123(5-6):229-35.
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.
马的精密切割肺切片（PCLS）用于评估几种药物化合物，即副交感神经阻滞剂阿托品、异丙托溴铵、噻托溴铵和磷酸二酯酶（PDE）4抑制剂西洛司特抑制乙酰甲胆碱（MCh）诱导的支气管收缩的能力。该研究使用了20匹马的新鲜肺样本，这些样本在安乐死和肺部检查后立即采集。取自头叶、尾叶和副叶的组织样本用于组织学检查。通过观察纤毛运动以及对添加不同浓度MCh（10^(-8)至10^(-4) mol/l）的支气管收缩反应，证实了250±20微米的PCLS的活力。在纯RPMI培养基中冲洗导致支气管重新张开。在含有西洛司特（10^(-5) mol/l）、阿托品、异丙托溴铵或噻托溴铵（10^(-7) mol/l）的RPMI溶液中孵育20分钟后，每组（随机分组，安慰剂对照）再次用MCh处理。重新张开后，通过添加MCh确认PCLS未受影响的活力。根据每次收缩的支气管横截面计算EC50值。添加阿托品、异丙托溴铵和噻托溴铵后，EC50值显著增加。西洛司特组与对照切片之间未发现差异。统计分析显示个体间差异大，但个体内差异小。临床检查、组织学发现与PCLS反应性之间未发现相关性。我们能够证实Vietmeier（2007年）关于马PCLS对MCh反应性的值，并使用一种新技术证明了几种新型支气管扩张剂的效果，该技术允许在同一匹马中评估多种物质。
