Kirsch C M, Sigal E, Djokic T D, Graf P D, Nadel J A
Cardiovascular Research Institute, University of California, San Francisco 94143-0130.
J Appl Physiol (1985). 1988 May;64(5):1792-5. doi: 10.1152/jappl.1988.64.5.1792.
We describe a new in vivo chemotaxis assay in the dog trachea using a double-balloon endotracheal catheter. When inflated, the two balloons isolate a segment of trachea, which is perfused through Silastic tubes using a peristaltic pump. After instilling a chemotactic agent, the perfusate is sampled periodically to permit characterization of the chemotactic response. We anesthetized four mongrel dogs and ventilated them mechanically through the double-balloon catheter. Two mediators, leukotriene B4 (LTB4) and 8S,15S-dihydroxyeicosatetraenoic acid (8,15-diHETE) were tested in each dog by perfusing the trachea with each mediator in Hanks' balanced salt solution (HBSS) containing ethanol and antibiotics. Aliquots were removed for differential cell counts at fixed time intervals over a 4-h period. Control experiments performed in each dog with the identical concentrations of ethanol and antibiotics in HBSS showed no cellular response before 180 min. At 240 min, the cell counts were 86 +/- 28 (SE) granulocytes/microliter (n = 4). In contrast, both LTB4 and 8,15-diHETE gave a significant cellular response at 120 min (309 +/- 125 and 141 +/- 41 granulocytes/microliter, respectively; P less than 0.05) but did not differ significantly from each other. These results suggest that both LTB4 and 8,15-diHETE can incite inflammatory responses in the dog trachea in vivo. Furthermore, the double-balloon catheter technique promises to be a useful in vivo chemotaxis assay.
我们描述了一种在犬气管中使用双气囊气管导管进行的新的体内趋化性测定方法。当气囊充气时，两个气囊隔离出一段气管，通过蠕动泵经硅橡胶管对其进行灌注。注入趋化剂后，定期采集灌注液以表征趋化反应。我们麻醉了4只杂种犬，并通过双气囊导管对其进行机械通气。在每只犬中，通过在含有乙醇和抗生素的汉克斯平衡盐溶液（HBSS）中用每种介质灌注气管，测试了两种介质，即白三烯B4（LTB4）和8S，15S-二羟基二十碳四烯酸（8,15-二氢二十碳四烯酸，8,15-diHETE）。在4小时内，每隔固定时间间隔取出等分试样进行细胞分类计数。在每只犬中用HBSS中相同浓度的乙醇和抗生素进行的对照实验显示，在180分钟之前没有细胞反应。在240分钟时，细胞计数为86±28（SE）个粒细胞/微升（n = 4）。相比之下，LTB4和8,15-diHETE在120分钟时均产生了显著的细胞反应（分别为309±125和141±41个粒细胞/微升；P<0.05），但彼此之间没有显著差异。这些结果表明，LTB4和8,15-diHETE均可在犬气管中引发体内炎症反应。此外，双气囊导管技术有望成为一种有用的体内趋化性测定方法。
