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我们建立了体外灌注内皮细胞单层研究血管通透性的方法,可以测定滤过系数(Kf)以及蛋白质渗透压反射系数(σ)。用Hanks平衡盐液(HBSS)或5 g/L白蛋白HBSS灌注致密内皮单层,Kf值分别是10.1±0.75和3.6±0.75μl·min~(-1)·cm~(-2)·kPa~1(n=3,±s)、说明白蛋白可降低内皮单层对水和小分子物质的通透性。血小板激活因子(PAF)10~8 mol/L作用30 min,用HBSS或白蛋白溶液灌注时Kf值分别增加到193.1％和133.3％。对照组蛋白清除率和σ分别是8.0±3.22μl·min~(-1)·cm~(-2)和0.37±0.09,PAF组分别是12.2±2.95μl·min~(-1)·cm~(-2)和0.18±0.06,相差显著,说明PAF可增加内皮单层的通透性。计算机图像分析表明,PAF作用后内皮细胞面积减小,形状因子、细胞间距增大,提示内皮细胞发生了收缩,这可能是PAF增加血管壁通透性的重要机制。
We established a method of perfusing vascular endothelial monolayer to study vascular permeability in vitro and determined the filtration coefficient (Kf) and the osmotic pressure reflection coefficient (σ). The dense endothelial monolayers were infused with Hanks balanced salt solution (HBSS) or 5 g / L albumin HBSS with Kf values of 10.1 ± 0.75 and 3.6 ± 0.75 μl · min -1 · cm -2 · kPa, respectively ~ 1 (n = 3, ± s), indicating that albumin reduces the permeability of monolayers to water and small molecules. Platelet activating factor (PAF) 10 ~ 8 mol / L for 30 min, with HBSS or albumin solution Kf value increased to 193.1% and 133.3%. The protein clearance rate and σ in the control group were 8.0 ± 3.22 μl · min -1 · cm -2 and 0.37 ± 0.09, respectively, and those in the PAF group were 12.2 ± 2.95 μl · min -1 · cm -1. (-2) and 0.18 ± 0.06, significant difference, indicating that PAF can increase the endothelial monolayer permeability. Computer image analysis showed that the area of endothelial cells decreased, the shape factor and the intercellular distance increased after PAF treatment, suggesting that endothelial cells contracted, which may be an important mechanism of PAF increasing vascular permeability.