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.
ABSTRACT
This study investigated the protective effect of two flavonols quercetin and myricetin on barrier function of rat intestinal epithelial (IEC-6) cells with indomethacin injury. When the cells were pretreated with the heated or unheated flavonols of 2.5-10 µmol/L for 24-48 h and then injured by 300 µmol/L indomethacin for 24 h, they showed reduced lactate dehydrogenase release (LDH) but increased cell viability; however, the flavonols of 20 µmol/L exerted a little effect to increase cell viability or decrease LDH release. Cell pretreatment with 5 µmol/L flavonols also resisted cell barrier dysfunction by increasing transepithelial resistance, reducing paracellular permeability, and promoting mRNA and protein expression of three tight junction proteins zonula occluden-1, occludin, and claudin-1. Although indomethacin injury increased intracellular Ca2+ concentration ([Ca2+]i) and consequently caused JNK/Src activation, the flavonols could decrease [Ca2+]i and attenuate the calcium-mediated JNK/Src activation. Quercetin with less hydroxyl groups was more efficient than myricetin to resist barrier dysfunction, while the unheated flavonols were more active than the heated counterparts to perform this effect. It is thus proposed that quercetin and myricetin could resist barrier dysfunction of the intestine once injured by indomethacin, but heat treatment of flavonols had a negative impact on barrier-protective function of flavonols.
