A dual-tracer method for differentiating transendothelial transport from paracellular leakage in vivo and in vitro.
A dual-tracer method for differentiating transendothelial transport from paracellular leakage in vivo and in vitro.
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DOI：
10.3389/fphys.2012.00166
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发表时间：
2012
影响因子：
4
通讯作者：
Lominadze D
中科院分区：
文献类型：
--
作者：
Muradashvili N;Tyagi R;Lominadze D
炎症引起的血管内皮功能受损可引起血浆蛋白渗漏，从而导致水肿。蛋白质可以通过两个主要的细胞旁通路和细胞外通路离开血管腔。由于前者涉及内皮细胞（EC）连接蛋白，后者涉及小泡形成，这两种途径是相互关联的。因此，在引起炎症的病理过程中，很难区分一种或另一种途径的主要作用。在这里，我们提出了一种新开发的双示踪探测方法，可以在病理过程中区分细胞间运输和细胞旁运输。这种基于荧光的方法可以在体外和体内检测各种动物血管制剂中EC层通透性的变化。该方法是基于生理和病理条件下低分子量分子（LMWM）与高分子量分子（HMWM）通过EC层或血管壁运输的比较。由于LMWM主要通过细胞旁渗漏，并且HMWM将通过细胞旁（当ECs之间的间隙足够宽时）和跨细胞途径移动，因此（在正常情况下和病理情况下）这些分子的运输速率差异将表明参与整个血管壁蛋白质穿越的主要运输途径。因此，通过活体显微镜评估不同大小示踪剂在体内的转运动力学的新方法可以澄清与微血管通透性升高相关的各种病理过程中药物递送靶标途径的识别相关的问题。
Inflammation-induced impaired function of vascular endothelium may cause leakage of plasma proteins that can lead to edema. Proteins may leave the vascular lumen through two main paracellular and transcellular pathways. As the first involves endothelial cell (EC) junction proteins and the second caveolae formation, these two pathways are interconnected. Therefore, it is difficult to differentiate the prevailing role of one or the other pathway during pathology that causes inflammation. Here we present a newly developed dual-tracer probing method that allows differentiation of transcellular from paracellular transport during pathology. This fluorescence-based method can be used in vitro to test changes in EC layer permeability and in vivo in various animal vascular preparations. The method is based on comparison of low molecular weight molecule (LMWM) transport to that of high molecular weight molecule (HMWM) transport through the EC layer or the vascular wall during physiological and pathological conditions. Since the LMWM will leak through mainly the paracellular and HMWM will move through paracellular (when gaps between the ECs are wide enough) and transcellular pathways, the difference in transport rate (during normal conditions and pathology) of these molecules will indicate the prevailing transport pathway involved in overall protein crossing of vascular wall. Thus, the novel approach of assessing the transport kinetics of different size tracers in vivo by intravital microscopy can clarify questions related to identification of target pathways for drug delivery during various pathologies associated with elevated microvascular permeability.
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影响因子：
8.3
作者：
del Zoppo GJ;Levy DE;Wasiewski WW;Pancioli AM;Demchuk AM;Trammel J;Demaerschalk BM;Kaste M;Albers GW;Ringelstein EB
通讯作者：
Ringelstein EB
影响因子：
5.5
作者：
CURRY, FE;MASON, JC;MICHEL, CC
通讯作者：
MICHEL, CC
DOI：
10.1161/01.atv.0000041037.06509.c2
发表时间：
2003-01-01
影响因子：
8.7
作者：
Gulledge, AA;McShea, C;Lord, ST
通讯作者：
Lord, ST
影响因子：
20.1
作者：
Hu, Guochang;Vogel, Stephen M.;Minshall, Richard D.
通讯作者：
Minshall, Richard D.
影响因子：
7.7
作者：
Bonnardel-Phu, E;Wautier, JL;Vicaut, E
通讯作者：
Vicaut, E