ALVEOLAR MACROPHAGE PARTICLE RELATIONSHIPS DURING LUNG CLEARANCE
ALVEOLAR MACROPHAGE PARTICLE RELATIONSHIPS DURING LUNG CLEARANCE
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DOI：
10.1165/ajrcmb/1.2.145
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发表时间：
1989-08-01
影响因子：
6.4
通讯作者：
TIETJEN, GL
中科院分区：
文献类型：
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作者：
LEHNERT, BE;VALDEZ, YE;TIETJEN, GL
材料和方法动物和颗粒滴注本研究使用成年雄性Fischer 344 只大鼠（250 至Z15 g，无特定病原体；Harlan Sprague-Dawley，印第安纳波利斯，印第安纳州）。将大鼠饲养在覆盖有纺丝聚酯过滤器（杜邦 22 号纺粘聚酯过滤器；EE DuPont, Co., Wilmington, DE）的悬浮笼中，并置于接受 HEPA 过滤空气的屏障设施中。一组大鼠在 Bthrane”（安氟烷；Airco，麦迪逊，威斯康星州）下气管内滴注 0.4 ml PBS（pH 7.3），如前所述（14）。另一组则滴注 0.4 ml PBS，其中含有 1.6 X 107（“-'86 ug）预洗（14）荧光、羧化、聚苯乙烯微球，其几何直径为2.13 11m（Fluoresbrite，羧化微球；Polysciences, Inc.，Warrington，PA）。滴注后，将动物垂直放置15至30秒，然后放回笼中。 支气管肺泡灌洗和无肺细胞通过腹膜内注射50mg戊巴比妥钠，在指定的滴注后时间开始处死大鼠。在呼吸暂停之前通过颈动脉横断对动物进行放血，并用用结扎线固定的钝的18号针对每只大鼠的气管插管。此后，气管和肺被整体切除，食道和心脏被切除。然后通过两个连续系列的肺清洗（灌洗系列1和灌洗系列2）来灌洗肺部。每个系列的共同点是，用 8 ml 室温 PBS 将肺部清洗六次，同时轻轻按摩。每个系列期间回收的液体被汇集在保持在冰中的单独管中。通常，在两次灌洗系列期间滴注的洗涤体积的> 94%被回收而没有肺渗漏。然而，当洗肺过程中发生过度渗漏时，即恢复了< 90%的滴注体积时，一些动物被排除在研究之外。用血细胞计数器对每个灌洗系列收获的细胞总数进行计数。细胞离心
Materials and MethodsAnimals and Particle Instillations Adult, male Fischer 344 rats (250 to Z15 g, specific pathogen-free; Harlan Sprague-Dawley, Indianapolis, IN) were used in this study. The rats were maintained in suspension cages covered with spun polyester filters (DuPont No. 22 spinbound polyester filter; EE DuPont, Co., Wilmington, DE) in a barriered facility that received HEPA filtered air. One group of rats was intratracheally instilled with 0.4 ml PBS (pH 7.3) under Bthrane"(enflurane; Airco, Madison, WI), as previously described (14). Another group was instilled with 0.4 ml PBS that contained 1.6 X 107 ("-'86 ug) prewashed (14) fluorescent, carboxylated, polystyrene microspheres that had a geometric diameter of 2.13 11m (Fluoresbrite, carboxylated microspheres; Polysciences, Inc., Warrington, PA). After the instillations, the animals were held vertically for 15 to 30 s before being returned to their cages.Bronchoalveolar Lavage and Lung Free Cells Rat killings at the indicated postinstillation times were initiated by intraperitoneal injections of 50 mg pentobarbital sodium. The animals were exsanguinated prior to apnea via carotid artery transection, and the trachea of each rat was cannulated with a blunt, 18-gauge needle secured with ligature. Thereafter, the trachea and lung were excised en bloc, and the esophagus and heart were removed. The lungs were then lavaged by two consecutive series of lung washings (lavage series 1 and lavage series 2). Common to each series, the lungs were washed six times with 8 ml of room-temperature PBS while they were gently massaged. Fluids retrieved during each series were pooled in separate tubes maintained in ice. Usually,> 94% of the wash volumes instilled during the two lavage series was recovered without lung leakage. However, some animals were excluded from the study when excessive leakage occurred during the lung washings, ie,< 90% of the instilled volume was recovered. The total numbers of cells harvested by each of the lavage series were counted with a hemocytometer. Cytocentrifuged