Bile Salts at Low pH Cause Dilation of Intercellular Spaces in In Vitro Stratified Primary Esophageal Cells, Possibly by Modulating Wnt Signaling

• Published in: Journal of gastrointestinal surgery 2016-03, Vol.20 (3), p.500-509
• Main Authors: Ghatak, Sayak, Reveiller, Marie, Toia, Liana, Ivanov, Andrei I., Zhou, Zhongren, Redmond, Eileen M., Godfrey, Tony E., Peters, Jeffrey H.
• Format: Article
• Language: English
• Subjects: Acids; beta Catenin - metabolism; Bile; Bile Acids and Salts - pharmacology; Biopsy; Cell Culture Techniques; Electric Impedance; Epithelial Cells - drug effects; Epithelium - drug effects; Epithelium - pathology; Esophagus; Esophagus - drug effects; Esophagus - pathology; Extracellular Space - drug effects; Gastroenterology; Gastroesophageal reflux; Humans; Medicine; Medicine & Public Health; Mucous Membrane - drug effects; Mucous Membrane - pathology; Original Article; Permeability; Polyesters; Signal Transduction - drug effects; Surgery; Telomerase
• ISSN: 1091-255X; 1873-4626
• DOI: 10.1007/s11605-015-3062-2

Background The presence of dilated intercellular spaces in the stratified squamous lining of the esophagus is the pathognomonic feature of reflux esophagitis secondary to gastroesophageal reflux disease (GERD). In addition to stomach acid, bile salts are major constituents of gastroesophageal refluxate. The aim of our study was to determine the effect of bile salts cocktail at different pHs on epithelial junctions in an in vitro transwell model of stratified esophageal squamous epithelium. Discussion Human telomerase reverse transcriptase (hTERT) immortalized primary esophageal EPC1 cells were grown on polyester transwell surfaces in calcium-enriched media. The cells exhibited gradual stratification into an 11-layered squamous epithelium over 7 days, together with epithelial barrier function as indicated by increased transepithelial electrical resistance (TEER). This stratified epithelium demonstrated well-formed tight junctions, adherens junctions, and desmosomes as visualized by immunofluorescence and electron microscopy. When exposed to short pulses of bile salts at pH 5, but not either condition alone, there was loss of stratification and decrease in TEER, concomitant with disruption of adherens junctions, tight junctions, and desmosomes, leading to the appearance of dilated intercellular spaces. At the cellular level, bile salts at pH 5 activated the Wnt pathway (indicated by increased β-catenin Ser552 phosphorylation). Conclusion In conclusion, in our in vitro transwell model bile salts at pH 5, but not bile salts or media at pH 5 alone, modulate Wnt signaling, disrupt different junctional complexes, and cause increased permeability of stratified squamous esophageal epithelium. These changes approximate the appearance of dilated intercellular space similar to that found in GERD patients.