to assess whether eupatilin affects H2O2 caused 5 LOX term in EECs western blotting analysis was conducted. After pre treatment with the indicated HSP inhibitor concentration of eupatilin for 12 hours, EECs were further exposed to 300 uM 316 Fig. 1. Influence of H2O2 on the cell viability of Effect and feline EECs of eupatilin on the H2O2 induced cell viability. Serum deprived EECs were incubated with H2O2 for twenty four hours at the indicated concentration. The cell viability was estimated using MTT assay. The morphologic changes of EECs were seen. Serumstarved EECs were incubated in the presence of eupatilin alone for 12 hours at the indicated concentration. the cells were incubated within the 600 uM H2O2 with or without eupatilin 12 hours before and throughout 24 hours, and then their survival was estimated utilizing the MTT assay and the morphologic alterations of cells were observed. Data are expressed as Means S. E of four experiments. Fig. 2. Ramifications of eupatilin to the H2O2 induced 5 LOX phrase. Serum deprived EECs were treated with H2O2 for 24 hours at each dose. Serum deprived cells were preincubated Posttranslational modification (PTM) in the existence of eupatilin for 12 hours at the indicated concentration and then stimulated with 300 uM H2O2 for 24 hours. 5 LOX expression was believed by Western blot. Data are expressed as Means S. E of three studies. H2O2 in the presence of eupatilin for 24-hours. Furthermore, pre-treatment with 150 uM eupatilin dramatically reduced the H2O2 induced 5 LOX protein expression. These indicated that p38 MAPK, JNK and ROS scavenging motion might mediate the inhibitory effect of eupatilin to the 5 LOX phrase by H2O2. These data were similar to the of the 5 LOX appearance by H2O2 with or without inhibitors. The phosphorylation of p38 MAPK and JNK was investigated, effect of H2O2 on activation of MAPKs To look for the influence of H2O2 on activation of MAPKs. The concentration dependence of p38 MAPK and JNK OSI-420 EGFR inhibitor phosphorylation was investigated by Western blot analysis. The change in the degree of phosphorylated p38 MAPK was estimated by Western blot analysis. The change of phosphorylated JNK level was estimated by Western blot analysis. The ROS scavengers offered similar effect to Eupatilin, and MAPK inhibitors showed further decrease down seriously to thirty days, similar to that of the non treated group. In this review, the addition of external H2O2 to esophageal epithelial cells exhibited significant cytotoxicity. The cell viability was decreased and the forms of cells were remarkably altered. But, eupatilin enhanced the reduced amount of cell viability by H2O2. Previously, we identified the cytoprotective properties of eupatilin might be caused by the induction of the antioxidant protein heme oxygenase 1 in ileal smooth muscle cells or esophageal epithelial cells. We also established that eupatilin induced HO 1 expression in esophageal epithelium of mice in vivo.