In vitro assessment of epithelial electrical resistance in human esophageal and jejunal mucosae and in Caco-2 cell layers
In vitro assessment of epithelial electrical resistance in human esophageal and jejunal mucosae and in Caco-2 cell layers
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DOI：
10.3109/00365521.2012.722677
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发表时间：
2012-11-01
影响因子：
1.9
通讯作者：
Fandriks, Lars
中科院分区：
文献类型：
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作者：
Bjorkman, Eleonora;Casselbrant, Anna;Fandriks, Lars
Objective.需要一种允许研究在不同临床条件下收集的人类粘膜标本的技术。这项研究阐明，如果方波脉冲分析区分上皮和跨壁电阻，如果有一个关联与分子探针的跨上皮渗透性。方法.在Ussing室中研究了食管（手术切除：n = 14;内镜活检：n = 15）和空肠（n = 12）的粘膜和Caco-2细胞单层。通过使用标准化的电流脉冲来记录上皮电阻。使用两种荧光素标记的探针（重量376和4000 Da）评估渗透性。结果食管粘膜的基线上皮电阻（类似于280 Ω *cm（2））高于空肠（类似于10 Ω *cm（2））和Caco-2细胞（类似于140 Ω *cm（2））。空肠标本（+88%）和Caco-2细胞（+75%）中上皮下对跨壁阻力的贡献高于食管（+30%）。在缺氧时，空肠粘膜和Caco-2细胞的上皮下电阻不变，而上皮电阻显著下降。这些发现与经上皮探针渗透性增加和形态学紊乱体征一致。食管上皮细胞耐缺氧。然而，暴露于脱氧胆酸和胰蛋白酶废除食管上皮电阻和增加探针渗透性。与健康受试者相比，糜烂性反流病患者的内镜食管活检显示出显著较低的上皮电阻和较高的电流。结论在Ussing室中的方波脉冲分析适用于评估上皮电阻，其可以反映具有已知尺寸的分子探针的跨上皮渗透性。此外，该技术区分健康和反流病食管粘膜活检。
Objective. There is a need for a technique allowing studies of human mucosal specimens collected during different clinical conditions. This study elucidates if square wave pulse analysis discriminates between epithelial and transmural electrical resistance and if there is an association with transepithelial permeability of molecular probes. Methods. Mucosae from esophagus (surgical resections: n = 14; endoscopic biopsies: n = 15) and jejunum (n = 12) and Caco-2 cell monolayers were investigated in Ussing chambers. Transmural and epithelial electrical resistance were recorded by the use of standardized current pulses. Permeability was assessed using two fluorescein-labeled probes (weight 376 and 4000 Da). Results. Baseline epithelial electrical resistance was higher in esophageal mucosa (similar to 280 Omega*cm(2)), than in jejunal (similar to 10 Omega*cm(2)) and Caco-2 cells (similar to 140 Omega*cm(2)). The subepithelial contribution to the transmural resistance was higher in jejunal preparations (+88%) and Caco-2 cells (+75%), than in esophageal (+30%). During hypoxia the subepithelial resistance was unchanged, whereas the epithelial resistance decreased significantly in jejunal mucosa and Caco-2 cells. These findings coincided with increased transepithelial probe permeability and signs of disturbed morphology. Esophageal epithelia were resistant to hypoxia. However, exposure to deoxycholic acid and trypsin abolished the esophageal epithelial resistance and increased probe permeability. Endoscopic esophageal biopsies from patients with erosive reflux disease exhibited significantly lower epithelial resistance and higher current than healthy subjects. Conclusion. Square wave pulse analysis in Ussing chambers is suitable for assessment of epithelial electrical resistance that can reflect transepithelial permeability of molecular probes with known size. Moreover, the technique discriminated between healthy and reflux-diseased esophageal mucosal biopsies.