Abstract：
Altered esophageal ion transport mechanisms play a key role in inflammatory and cancerous diseases of the esophagus, but epithelial ion processes have been less studied in the esophagus because of the lack of a suitable experimental model. In this study, we generated three-dimensional (3D) esophageal organoids (EOs) from two different mouse strains and characterized the ion transport processes of the EOs. EOs form a cell-filled structure with a diameter of 250-300 µm and were generated from epithelial stem cells as shown by FACS analysis. Using conventional PCR and immunostaining, the presence of Slc26a6 Cl-/HCO3- anion exchanger (AE), Na+/H+ exchanger (NHE), Na+/HCO3- cotransporter (NBC), cystic fibrosis transmembrane conductance regulator (CFTR), and anoctamin 1 Cl- channels was detected in EOs. Microfluorimetric techniques revealed high NHE, AE, and NBC activities, whereas that of CFTR was relatively low. In addition, inhibition of CFTR led to functional interactions between the major acid-base transporters and CFTR. We conclude that EOs provide a relevant and suitable model system for studying the ion transport mechanisms of esophageal epithelial cells, and they can be also used as preclinical tools to assess the effectiveness of novel therapeutic compounds in esophageal diseases associated with altered ion transport processes.
摘要：
食管离子转运机制的改变在食管炎症和癌性疾病中起着关键作用。但是由于缺乏合适的实验模型，在食道中对上皮离子过程的研究较少。在这项研究中,我们从两种不同的小鼠品系中生成了三维（3D）食管类器官（EO），并表征了EO的离子转运过程。EO形成直径为250-300μm的细胞填充结构，并由上皮干细胞产生，如FACS分析所示。使用常规PCR和免疫染色，Slc26a6Cl-/HCO3-阴离子交换剂（AE）的存在，Na+/H+交换剂(NHE),Na+/HCO3-共转运蛋白(NBC),囊性纤维化跨膜传导调节因子（CFTR），并且在EO中检测到anocamin1Cl-通道。微荧光技术显示高NHE，AE,和NBC的活动，而CFTR相对较低。此外,CFTR的抑制导致主要酸碱转运蛋白和CFTR之间的功能相互作用。我们得出结论，EOs为研究食管上皮细胞的离子转运机制提供了相关且合适的模型系统。它们还可以用作临床前工具，以评估新型治疗化合物在与离子转运过程改变相关的食管疾病中的有效性。
