Objective To establish the injury model of human gastric mucosal epithelial cells(GES-1 cells)with acetic acid,and to explore the protective effect and mechanism of the alkaloid components Laetanine and Launobine from Lindera reflexa Hemsl.on the injury of GES-1 cells.Methods The optimal concentration of Laetanine and Launobine were determined by MTT cell proliferation/toxicity experiment.The culture medium containing 0.01%～0.2%acetic acid was used to act on GES-1 cells for 3,4 and 5 hours,respectively.The best modeling conditions were screened.The cell survival rate of GES-1 cells treated with Laetanine and Launobine was determined by acetic acid-induced cell injury model.The concentration of NO in cell supernatant was measured by Griess method.The levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and prostaglandin E2(PGE2)secreted in cell supernatant were measured by ELISA.The inhibitory rate of superoxide dismutase(SOD)was measured by WST-1,and the activity value of SOD was calculated.Results The treatment with 0.1%acetic acid solution for 3h was the best modeling condition for GES-1 cells injury.Compared with the model group,both Laetanine and Launobine could significantly increase the survival rate of GES-1 cells(P<0.01).After treatment with Laetanine and Launobine,the levels of NO,TNF-α,IL-6 and PGE2 in cell supernatant were significantly decreased,while the activity of SOD was significantly increased(P<0.01 or P<0.05).Conclusion The alkaloid components Laetanine and Launobine from Lindera reflexa Hemsl.can both protect GES-1 cells injured by acetic acid,reduce the degree of cells damage.The mechanism of action may be related to the decrease of NO,TNF-α,IL-6 and PGE2 levels and the increase of SOD level.These results indicate that Laetanine and Launobine may be effective components of Lindera reflexa Hemsl.for anti-gastric ulcer.
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