The protective mechanism of QGC in feline esophageal epithelial cells by interleukin-1 beta treatment

Abstract

In a previous study, Quercetin-3-O-beta-d-glucuronopyranoside (QGC) has anti-oxidative and anti-inflammatory effects in vivo. QGC is a flavonoid glucoside extracted from Rumex Aquaticus. We investigated the downstream target proteins involved in IL-1 beta-stimulated ROS production and the ability of QGC to inhibit ROS production. Cell viability was determined using the MTT reduction assay. Western blot analysis was performed with antibodies to investigate the activation of three MAPKs, NF-kappa B, and phosphorylated I kappa B-alpha (pIB), and the expression of COX-2. 2',7'-dichlorofluorescin diacetate was used to detect the generation of intracellular ROS species. When the cells were exposed to media containing IL-1 beta for 18 h, cell viability was not affected. QGC did not reduce the COX-2 expression induced by IL-1 beta. However; QGC attenuated the production of intracellular ROS induced by IL-1 beta. IL-1 beta increased the expression of ERK, p38 MAPK, and pIB, and nuclear translocation of NF-kappa B were recovered by the ROS scavenger N-acetyl-l-cysteine (NAC) and QGC, but not by the NADPH oxidase inhibitor diphenylene iodonium. Pretreatment of cells with the ERK inhibitor PD98059, the p38 MAPK inhibitor SB202190, NAC, and QGC attenuated nuclear translocation of NF-kappa B and activation of pIB. QGC has a scavenging effect on cytokine-induced ROS production, thereby preventing its downstream effects, nuclear translocation of NF-kappa B, and activation of pIB is mediated by activation of ERK and p38 MAPK, although QGC does not inhibit IL-1 beta-stimulated COX-2 expression in feline esophageal epithelial cells. The data suggest that QGC exerts anti-oxidative effects and inhibitory effects against esophageal epithelial cells signals by the action of IL-1 beta treatment.