Gordon Sharon A, Lominadze David, Saari Jack T, Lentsch Alex B, Schuschke Dale A
Department of Physiology and Biophysics, Health Sciences Center A1115, University of Louisville, Louisville, KY 40292, USA.
Exp Biol Med (Maywood). 2005 Sep;230(8):543-8. doi: 10.1177/153537020523000805.
We have previously shown that dietary copper deficiency augments neutrophil accumulation in the lung microvasculature. The current study was designed to determine whether a diet deficient in copper promotes neutrophil chemoattraction within the lung vasculature or if it alters the mechanical properties of the neutrophil, thus restricting passage through the microvessels. Sprague-Dawley rats were fed purified diets that were either copper adequate (6.3 microg Cu/g diet) or copper deficient (0.3 microg Cu/g diet) for 4 weeks. To assess neutrophil chemoattraction, bronchoalveolar lavage fluid was assayed for the neutrophil chemokine macrophage inflammatory protein-2 (MIP-2) by enzyme-linked immunosorbent assay. Neutrophil deformability was determined by measuring the pressure required to pass isolated neutrophils through a 5-microm polycarbonate filter. The MIP-2 concentration was not significantly different between the dietary groups (Cu adequate, 435.4 +/- 11.9 pg/ml; Cu deficient, 425.6 +/- 14.8 pg/ml). However, compared with controls, more pressure was needed to push Cu-deficient neutrophils through the filter (Cu adequate, 0.150 +/- 0.032 mm Hg/sec; Cu deficient, 0.284 +/- 0.037 mm Hg/sec). Staining of the filamentous actin (F-actin) with FITC-Phalloidin showed greater F-actin polymerization and shape change in the Cu-deficient group. These results suggest that dietary copper deficiency reduces the deformability of neutrophils by promoting F-actin polymerization. Because most neutrophils must deform during passage from arterioles to venules in the lungs, we propose that copper-deficient neutrophils accumulate in the lung because they are less deformable.
我们之前已经表明,饮食中铜缺乏会增加肺微血管中中性粒细胞的积聚。当前的研究旨在确定铜缺乏饮食是否会促进肺血管内中性粒细胞的趋化作用,或者它是否会改变中性粒细胞的机械特性,从而限制其通过微血管。将Sprague-Dawley大鼠喂食纯化饮食4周,饮食分为铜充足组(6.3微克铜/克饮食)或铜缺乏组(0.3微克铜/克饮食)。为了评估中性粒细胞趋化作用,通过酶联免疫吸附测定法检测支气管肺泡灌洗液中的中性粒细胞趋化因子巨噬细胞炎性蛋白-2(MIP-2)。通过测量使分离的中性粒细胞通过5微米聚碳酸酯滤膜所需的压力来确定中性粒细胞的变形能力。饮食组之间的MIP-2浓度没有显著差异(铜充足组,435.4±11.9皮克/毫升;铜缺乏组,425.6±14.8皮克/毫升)。然而,与对照组相比,推动铜缺乏的中性粒细胞通过滤膜需要更大的压力(铜充足组,0.150±0.032毫米汞柱/秒;铜缺乏组,0.284±0.037毫米汞柱/秒)。用异硫氰酸荧光素-鬼笔环肽对丝状肌动蛋白(F-肌动蛋白)进行染色显示,铜缺乏组中F-肌动蛋白的聚合和形状变化更大。这些结果表明,饮食中铜缺乏通过促进F-肌动蛋白聚合降低了中性粒细胞的变形能力。由于大多数中性粒细胞在从肺中的小动脉进入小静脉的过程中必须变形,我们推测铜缺乏的中性粒细胞在肺中积聚是因为它们的变形能力较差。