一种增强肠道上皮屏障完整性和加速修复的新型药理学方法。
A Novel Pharmacological Approach to Enhance the Integrity and Accelerate Restitution of the Intestinal Epithelial Barrier.
机构信息
Department of Inflammation and Immunity, Lerner Research Institute of Cleveland Clinic Foundation, Cleveland, OH.
School of Nursing, Virginia Commonwealth University, Richmond, VA.
出版信息
Inflamm Bowel Dis. 2020 Aug 20;26(9):1340-1352. doi: 10.1093/ibd/izaa063.
BACKGROUND
Disruption of the gut barrier is an essential mechanism of inflammatory bowel diseases (IBDs) contributing to the development of mucosal inflammation. A hallmark of barrier disruption is the disassembly of epithelial adherens junctions (AJs) driven by decreased expression of a major AJ protein, E-cadherin. A group of isoxazole compounds, such as E-cadherin-upregulator (ECU) and ML327, were previously shown to stimulate E-cadherin expression in poorly differentiated human cancer cells. This study was designed to examine whether these isoxazole compounds can enhance and protect model intestinal epithelial barriers in vitro.
METHODS
The study was conducted using T84, SK-CO15, and HT-29 human colonic epithelial cell monolayers. Disruption of the epithelial barrier was induced by pro-inflammatory cytokines, tumor necrosis factor-α, and interferon-γ. Barrier integrity and epithelial junction assembly was examined using different permeability assays, immunofluorescence labeling, and confocal microscopy. Epithelial restitution was analyzed using a scratch wound healing assay.
RESULTS
E-cadherin-upregulator and ML327 treatment of intestinal epithelial cell monolayers resulted in several barrier-protective effects, including reduced steady-state epithelial permeability, inhibition of cytokine-induced barrier disruption and junction disassembly, and acceleration of epithelial wound healing. Surprisingly, these effects were not due to upregulation of E-cadherin expression but were mediated by multiple mechanisms including inhibition of junction protein endocytosis, attenuation of cytokine-induced apoptosis, and activation of promigratory Src and AKT signaling.
CONCLUSIONS
Our data highlight ECU and ML327 as promising compounds for developing new therapeutic strategies to protect the integrity and accelerate the restitution of the intestinal epithelial barrier in IBD and other inflammatory disorders.
背景
肠道屏障的破坏是炎症性肠病（IBD）的一个重要机制，导致黏膜炎症的发展。屏障破坏的一个标志是上皮细胞黏附连接（AJs）的解体，这是由主要 AJ 蛋白 E-钙黏蛋白表达减少驱动的。一组异噁唑化合物，如 E-钙黏蛋白上调剂（ECU）和 ML327，以前被证明可以刺激低分化人类癌细胞中 E-钙黏蛋白的表达。本研究旨在研究这些异噁唑化合物是否可以增强和保护体外模型肠道上皮屏障。
方法
该研究使用 T84、SK-CO15 和 HT-29 人结肠上皮细胞单层进行。通过促炎细胞因子肿瘤坏死因子-α和干扰素-γ诱导上皮屏障破坏。使用不同的通透性测定、免疫荧光标记和共聚焦显微镜检查来检测上皮屏障的完整性和上皮连接的组装。使用划痕愈合实验分析上皮修复。
结果
E-钙黏蛋白上调剂和 ML327 处理肠上皮细胞单层可产生多种屏障保护作用，包括降低稳态上皮通透性、抑制细胞因子诱导的屏障破坏和连接解体以及加速上皮伤口愈合。令人惊讶的是，这些作用不是由于 E-钙黏蛋白表达的上调，而是通过多种机制介导的，包括抑制连接蛋白内吞作用、减弱细胞因子诱导的细胞凋亡以及激活促迁移Src 和 AKT 信号。
结论
我们的数据强调了 ECU 和 ML327 作为有前途的化合物，可用于开发新的治疗策略，以保护 IBD 和其他炎症性疾病中肠道上皮屏障的完整性并加速其修复。
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