Galangin and Kaempferol Alleviate the Indomethacin-Caused Cytotoxicity and Barrier Loss in Rat Intestinal Epithelial (IEC-6) Cells Via Mediating JNK/Src Activation.

Abstract

Nonsteroidal anti-inflammatory drugs (NSAIDs) like indomethacin and others are widely used in clinics, but they have the potential to cause severe gastrointestinal damage including intestinal barrier dysfunction. Thus, two flavonols galangin and kaempferol with or without heat treatment (100 °C, 30 min) were assessed for their effect on indomethacin-damaged rat intestine epithelial (IEC-6) cells. In total, the cell exposure of 300 μmol/L indomethacin for 24 h caused cell toxicity efficiently, resulting in decreased cell viability, enhanced lactate dehydrogenase (LDH) release or reactive oxygen species (ROS) production, and obvious barrier loss. Meanwhile, pretreatment of the cells with these flavonols for 24 and 48 h before the indomethacin exposure could alleviate cytotoxicity and especially barrier loss, resulting in increased cell viability and transepithelial resistance, decreased LDH release, ROS production, and paracellular permeability, together with the promoted expression of three tight junction proteins zonula occluden-1, occludin, and claudin-1. Moreover, the intracellular Ca2+ concentration and expression levels of p-JNK and p-Src arisen from the indomethacin damage were also reduced by the flavonols, suggesting an inhibited calcium-mediated JNK/Src activation. Consistently, galangin showed higher activity than kaempferol to the cells, while the heated flavonols were less efficient than the unheated counterparts. It is thus highlighted that the two flavonols could alleviate indomethacin cytotoxicity and combat against the indomethacin-induced barrier loss in IEC-6 cells, but heat treatment of the flavonols would weaken the two beneficial functions.