In Vitro and In Vivo Approaches to Determine Intestinal Epithelial Cell Permeability
In Vitro and In Vivo Approaches to Determine Intestinal Epithelial Cell Permeability
复制标题
确定肠上皮细胞渗透性的体外和体内方法
DOI：
10.3791/57032
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发表时间：
2018-10-01
影响因子：
1.2
通讯作者：
He, Wei-Qi
中科院分区：
文献类型：
--
作者：
Li, Ban-Ruo;Wu, Jia;He, Wei-Qi
肠道屏障防御病原微生物和微生物毒素。其功能受紧密连接通透性和上皮细胞完整性的调节，肠屏障功能的破坏有助于胃肠道和全身性疾病的进展。本文介绍了两种简单的方法来测定肠上皮细胞的通透性。在体外，将Caco-2BBe细胞作为单层接种在组织培养威尔斯孔中，并且可以通过上皮（伏特/欧姆）计测量跨上皮电阻（TER）。该方法操作简便，重复性好，具有较强的可信度。在体内，用4kDa异硫氰酸荧光素（FITC）-葡聚糖管饲小鼠，并在从小鼠收集的血清样品中测量FITC-葡聚糖浓度以确定上皮渗透性。经口灌胃提供了准确的剂量，因此是体内测量肠道通透性的首选方法。这两种方法结合在一起，可以在体外和体内测量肠上皮的通透性，从而用于研究疾病与屏障功能之间的联系。
The intestinal barrier defends against pathogenic microorganism and microbial toxin. Its function is regulated by tight junction permeability and epithelial cell integrity, and disruption of the intestinal barrier function contributes to progression of gastrointestinal and systemic disease. Two simple methods are described here to measure the permeability of intestinal epithelium. In vitro, Caco-2BBe cells are plated in tissue culture wells as a monolayer and transepithelial electrical resistance (TER) can be measured by an epithelial (volt/ohm) meter. This method is convincing because of its user-friendly operation and repeatability. In vivo, mice are gavaged with 4 kDa fluorescein isothiocyanate (FITC)-dextran, and the FITC-dextran concentrations are measured in collected serum samples from mice to determine the epithelial permeability. Oral gavage provides an accurate dose, and therefore is the preferred method to measure the intestinal permeability in vivo. Taken together, these two methods can measure the permeability of the intestinal epithelium in vitro and in vivo, and hence be used to study the connection between diseases and barrier function.