DeFouw D O, Brown K L, Feinberg R N
Department of Anatomy, Cell Biology and Injury Sciences, UMDNJ-New Jersey Medical School, Newark 07103-2757.
J Vasc Res. 1993 May-Jun;30(3):154-60. doi: 10.1159/000158990.
Cultured canine jugular vein endothelial cells were seeded on polycarbonate filters to create an in vitro permeability assay. The calculated diffusive permeability coefficient for FITC-BSA across untreated monolayers was 1.1 +/- 0.4 x 10(-6) cm/s. After 15-min incubations with either histamine or bradykinin, the resistance to albumin flux across the monolayers was reduced significantly. Diffusive albumin permeability coefficients were 3.4 +/- 1.8 x 10(-6) and 4.1 +/- 2.0 x 10(-6) cm/s, respectively. Ultrastructural morphometric analyses of the endothelial cell monolayers served to define uniform dimensions of intercellular clefts and similar plasmalemmal vesicle densities in the untreated and the vasomediator-activated monolayers. These results are consistent with the interpretation that the vasomediator-activated pathway across the venous endothelial monolayers is not dependent on sustained intercellular gap formation or sustained expansion of the plasmalemmal vesicle population for the 15-min observation periods. Whether the increased albumin flux is dependent on transient gap formation or on physical changes within the venous endothelial cell glycocalyx remains to be tested.
将培养的犬颈静脉内皮细胞接种在聚碳酸酯滤膜上,以建立体外通透性测定法。计算得出,异硫氰酸荧光素标记的牛血清白蛋白(FITC-BSA)在未处理单层细胞上的扩散渗透系数为1.1±0.4×10⁻⁶ cm/s。在用组胺或缓激肽孵育15分钟后,单层细胞对白蛋白通量的阻力显著降低。扩散白蛋白渗透系数分别为3.4±1.8×10⁻⁶和4.1±2.0×10⁻⁶ cm/s。对内皮细胞单层进行超微结构形态计量分析,以确定未处理和血管介质激活的单层细胞中细胞间缝隙的均匀尺寸以及类似的质膜小泡密度。这些结果与以下解释一致:在15分钟的观察期内,血管介质激活的穿过静脉内皮单层细胞的途径不依赖于持续的细胞间间隙形成或质膜小泡数量的持续增加。白蛋白通量增加是否依赖于短暂的间隙形成或静脉内皮细胞糖萼内的物理变化仍有待测试。