Neutrophil-endothelial interactions in a cell-column model of the microvasculature: Effects of fMLP
Neutrophil-endothelial interactions in a cell-column model of the microvasculature: Effects of fMLP
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DOI：
10.3109/10739689609148306
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发表时间：
1996-09-01
期刊：
影响因子：
--
通讯作者：
Alexander, J. S.
中科院分区：
文献类型：
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作者：
Haselton, F. R.;Woodall, J. H.;Alexander, J. S.
目的：本研究的目的是开发一种体外模型，与停滞的两腔系统相比，该模型更类似于脉管系统中的流变条件，并且允许几乎同时测量多形核白细胞（PMN）截留和内皮单层通透性。方法：用 2 X 106 PMN/ml 灌注先前描述的脉管系统细胞柱模型，平均流体速度为 0.09 cm/s，平均估计内皮表面剪切应力为 4.5 达因/cm2，总共灌注 60 分钟。 PMN 截留量是根据该再循环系统中的 PMN 计数来估计的。通过对三种共注射染料的指示剂稀释分析来估计牛内皮渗透性：蓝色葡聚糖（分子量（MW）2 X 106）、荧光素钠（MW 342）和氰钴胺（MW 1355）。结果：尽管前 15 分钟内每 65 cm2 内皮表面循环 PMN 的数量以 14% 的速度减少，但通过细胞柱的 PMN 循环并未激活 PMN。内皮细胞单层通透性在60分钟内没有变化。添加甲酰基-甲硫氨酰-亮氨酰-苯丙氨酸 (fMLP) (10-5 M) 可激活 PMN，并在前 15 分钟内将循环 PMN 的截留率显着增加至每 65 cm2 的 27%。尽管捕获的循环 PMN 百分比增加，但向细胞柱灌注液中添加 10-5 M fMLP 或 PMN + fMLP 并未增加内皮单层通透性。结论：在 60 分钟的研究期间，即使在用 fMLP 刺激 PMN 后，微血管再循环模型中 PMN 的截留也与内皮通透性增加无关。
Objective: The purpose of this study was to develop an in vitro model which, compared to stagnant two-chamber systems, is more analogous to the rheological conditions found in the vasculature and permits near simultaneous measurement of polymorphonuclear leukocytes (PMN) entrapment and endothelial monolayer permeability. Methods: A previously described cell-column model of the vasculature was perfused with 2 X 106 PMN/ml at an average fluid velocity of 0.09 cm/s and an average estimated endothelial surface shear stress of 4.5 dyne/cm2 for a total of 60 min. PMN entrapment was estimated from counts of PMN in this recirculating system. Bovine endothelial permeability was estimated from an indicator dilution analysis of the three co-injected dyes: blue dextran (molecular weight (MW)) 2 X 106), sodium fluorescein (MW 342) and cyanocobalamin (MW 1355). Results: Circulation of PMN through cell-columns did not activate PMN, although the number of circulating PMN decreased at a rate of 14% per 65 cm2 of endothelial surface in the first 15 min. Endothelial cell monolayer permeability did not change within 60 min. Addition of formyl-methionyl-leucyl-phenylalanine (fMLP) (10-5 M) activated PMN and significantly increased entrapment of circulating PMN to 27% per 65 cm2 in the first 15 min. Although the percentage of circulating PMN entrapped increased, endothelial monolayer permeability was not increased by either the addition of 10-5 M fMLP or PMN + fMLP to the cell-column perfusate. Conclusions: Over the 60-min period studied, entrapment of PMN within a recirculating model of the microvasculature was not associated with an increase in endothelial permeability even after PMN stimulation with fMLP.