The Protective Effect of Eupatilin against Hydrogen Peroxide-Induced Injury Involving 5-Lipoxygenase in Feline Esophageal Epithelial Cells

The Protective Effect of Eupatilin against Hydrogen Peroxide-Induced Injury Involving 5-Lipoxygenase in Feline Esophageal Epithelial Cells

In this study, we focused to identify whether eupatilin (5,7-dihydroxy-3 ,4 ,6-trimethoxyflavone), an extract from Artemisia argyi folium, prevents H₂O₂-induced injury of cultured feline esophageal epithelial cells. Cell viability was measured by the conventional MTT reduction assay. Western blot analysis was performed to investigate the expression of 5-lipoxygenase by H₂O₂ treatment in the absence and presence of inhibitors. When cells were exposed to 600 ՌM H₂O₂ for 24 hours, cell viability was decreased to 40%. However, when cells were pretreated with 25∼150 ՌM eupatilin for 12 hours, viability was significantly restored in a concentration-dependent manner. H₂O₂-treated cells were shown to express 5-lipoxygenase, whereas the cells pretreated with eupatilin exhibited reduction in the expression of 5-lipoxygenase. The H₂O₂-induced increase of 5-lipoxygenase expression was prevented by SB202190, SP600125, or NAC. We further demonstrated that the level of leukotriene B₄ (LTB₄) was also reduced by eupatilin, SB202190, SP600125, NAC, or nordihydroguaiaretic acid (a lipoxygenase inhibitor) pretreatment. H₂O₂ induced the activation of p38MAPK and JNK, this activation was inhibited by eupatilin. These results indicate that eupatilin may reduce H₂O₂-induced cytotoxicity, and 5-lipoxygenase expression and LTB₄ production by controlling the p38 MAPK and JNK signaling pathways through antioxidative action in feline esophageal epithelial cells.