Fructooligosaccharides induce tight junction assembly in intestinal epithelial cells via AMPK activation

Abstract

Several intestinal and metabolic diseases including inflammatory bowel diseases (IBD), malnutrition, necrotizing enterocolitis and diabetes mellitus (DM) involve impairment of intestinal tight junction assembly. AMP‐activating protein kinase (AMPK) is an energy sensor that enhances tight junction assembly. Fructooligosaccharides (FOS) are well known prebiotics protecting against intestinal and metabolic diseases with unclear mechanisms. This study aimed to investigates effect of FOS on AMPK and tight junction assembly in intestinal epithelial cells (IEC). Using western blot analyses, it was found that FOS with concentrations from 0.001 to 10 mg/ml induced AMPK activation on T84 cells. FOS at concentration of 0.1 mg/ml showed the greatest effect on AMPK activation and it was used in the subsequent experiments. Calcium‐switch assays and fluorescein isothiocyanate (FITC) leakage experiments were then performed. The results revealed that FOS significantly increased percentage of TER at 15 minute, 1 hour, 6 hour, 12 hour and 24 hour post‐treatment. Interestingly, STO‐609, a CaMKKβ inhibitor, suppressed the effect of FOS on increasing TER and reducing FITC leakage. These results indicate that FOS increased tight junction assembly via CaMKKβ‐AMPK activation.Support or Funding InformationThis work is supported by Tonkla Ramathibodi program and Thailand Research Fund and Mahidol University (BRG5980008)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.