Anderson B O, Brown J M, Shanley P F, Bensard D D, Harken A H
Department of Surgery, University of Colorado Health Sciences Center, Denver 80262.
Surgery. 1991 Jan;109(1):51-61.
Neutrophils have been implicated in the pathogenesis of several inflammatory lung disorders including asthma, emphysema, and adult respiratory distress syndrome. The precursors of these destructive cells are thought to be marginating neutrophils that, although intravascular, remain intimately associated with endothelium, resisting the shearing forces of flowing blood. The purposes of this study were (1) to examine a method for quantitating marginating lung neutrophils, (2) to assess the adherence of marginating neutrophils to normal lung endothelium, and (3) to determine the reversibility of neutrophil-endothelial cell adherence. Rats were anesthetized and ventilated, and their lungs were exposed through a median sternotomy. The left lung was tied off with blood in situ and the right lung was perfused intraarterially with colloid for 2 minutes. In separate experiments, both left and right lungs were perfused intraarterially with colloid for 25 minutes. Myeloperoxidase, a neutrophil granule enzyme, was related to lung neutrophils. Dithionite-sensitive optical density (DSOD) was related to lung hemoglobin. Marginating lung neutrophils were quantitated by measuring the peroxidase activity of normal blood-perfused lung (myeloperoxidase assay) and subtracting from it the fraction of activity corresponding to the lung blood content (DSOD). The marginating neutrophil pool was identified by 2.1 units myeloperoxidase (5 x 10(6) neutrophils) per gm wet lung. Although the marginating pool was depleted by 54% during 2 minutes of lung colloid perfusion, lung erythrocytes (DSOD) were decreased by a significantly greater 93% (p less than 0.05). Lungs perfused with colloid for 25 minutes had negligible remaining myeloperoxidase activity. We conclude that (1) marginating lung neutrophils can be quantitated with the myeloperoxidase and DSOD assays, (2) marginating neutrophils are relatively adherent to normal lung endothelium compared with intravascular erythrocytes, and (3) the adherence of marginating neutrophils is fully reversible.
中性粒细胞与包括哮喘、肺气肿和成人呼吸窘迫综合征在内的多种炎症性肺部疾病的发病机制有关。这些具有破坏性的细胞的前体被认为是边缘性中性粒细胞,它们虽然位于血管内,但仍与内皮细胞紧密相连,抵抗流动血液的剪切力。本研究的目的是:(1)研究一种定量边缘性肺中性粒细胞的方法;(2)评估边缘性中性粒细胞对正常肺内皮细胞的黏附性;(3)确定中性粒细胞与内皮细胞黏附的可逆性。将大鼠麻醉并进行通气,通过正中胸骨切开术暴露其肺部。左肺原位结扎保留血液,右肺经动脉灌注胶体2分钟。在单独的实验中,左、右肺均经动脉灌注胶体25分钟。髓过氧化物酶是一种中性粒细胞颗粒酶,与肺中性粒细胞相关。连二亚硫酸盐敏感光密度(DSOD)与肺血红蛋白相关。通过测量正常血液灌注肺的过氧化物酶活性(髓过氧化物酶测定),并从中减去与肺血含量相对应的活性部分(DSOD),对边缘性肺中性粒细胞进行定量。边缘性中性粒细胞池通过每克湿肺2.1单位髓过氧化物酶(5×10⁶个中性粒细胞)来确定。尽管在肺胶体灌注2分钟期间边缘性中性粒细胞池减少了54%,但肺红细胞(DSOD)减少了显著更多的93%(p<0.05)。灌注胶体25分钟的肺髓过氧化物酶活性可忽略不计。我们得出结论:(1)可通过髓过氧化物酶和DSOD测定对边缘性肺中性粒细胞进行定量;(2)与血管内红细胞相比,边缘性中性粒细胞相对黏附于正常肺内皮细胞;(3)边缘性中性粒细胞的黏附是完全可逆的。