Fructose Promotes Oxidative Stress, Sphingolipid Remodelling and Transient Intestinal Barrier Dysfunction in Differentiated Caco-2 Cells

Background/Objectives: High dietary fructose consumption has been linked to impaired intestinal barrier function and several metabolic disorders. Although oxidative stress has been proposed as a mediator of fructose-related epithelial injury, the involvement of tight-junction (TJ) proteins and sphingolipid metabolism is still not fully defined. Here, differentiated Caco-2 monolayers were exposed to fructose (1–50 mM) to examine changes in epithelial barrier integrity and associated molecular responses.

Methods

Cell viability, transepithelial electrical resistance (TEER), reactive oxygen species (ROS), intracellular reduced and oxidized glutathione (GSH/GSSG) ratio, nitrites as an indirect assessment of nitric oxide (NO) production, TJ protein level (ZO-1, JAM-A and occludin), sphingolipid profile by UHPLC-QTOF-MS, and expression of the ceramide synthases CERS2 and CERS6 were evaluated.