Protective Effects And Its Mechanism Of Rebamipide On Aspirin-induced Injury In Human Gastric Mucosal Epithelium Cells

Objective:To investigate the protective effects and its mechanism of rebamipide on aspirin-induced injury in human gastric mucosal epithelium cells (GES-1)Methods:GES-1cells monolayer culture model was established in vitro. Then the cells were divided into negative control group, aspirin injury group and combination of rebamipide at different concentration (0.2,0.5,1.0mmol/L) and aspirin groups. After24h incubation, the cell proliferation, the content of malondialdehyde (MDA), the activity of superoxide dismutase (SOD) and heme oxygenase-1(HO-1) of each group were detected respectively. The ultrastructural changes of each group were observed by transmission electron microscopy (TEM). The expressions of nuclear factor erythroid2-related factor2(Nrf2) and HO-1at protein level in the cells of each group were detected by Western blot. Nrf2interfering suppression test was performed and the influence of Nrf2small interfering RNA (siRNA) on the expression of HO-1protein was observed.Results:The MTT results showed that the cell viability of aspirin injured group and combination of rebamipide at different concentration (0.2,0.5,1.0mM) and aspirin groups were (49.56±3.88)%,(59.34±4.36)%,(70.79±5.96)%and (86.07±5.20)%, respectively (F=30.634, P<0.01). Compared with aspirin injured group, the content of MDA was significantly lowered in combination of rebamipride at different concentration (0.2,0.5,1.0mM) and aspirin groups (F=23.821, P<0.05). Compared with aspirin injured group, the activity of SOD was significantly increased in combination of rebamipide at0.5and1.0mmol/L and aspirin groups (F=25.666, P<0.05), but there was no statistical significance between aspirin injured group and combination of rebamipide at0.2mM and aspirin group (t=1.586, P>0.05). Under TEM, the cell ultrastructural was obviously injured after aspirin treated, while rebamipide could relieve the injury. Compared with aspirin injured group, the activity of HO-1was significantly increased in combination of rebamipide at different concentration (0.2,0.5,1.0mM) and aspirin groups (F=50.055, P<0.05). The Western blot analysis showed that expressions of Nrf2and HO-1at protein level among combination of rebamipide at0.2,0.5,1.0mmol/L and aspirin groups were all significantly up-regulated compared with that in aspirin injured group (F=92.550、38.235, both P<0.05). After transfected with Nrf2siRNA, the expression of HO-1in aspirin injured group and combination of rebamipide and aspirin group was significantly lower than that before transfection (t=6.276、10.444, both P<0.05).Conclusion:Aspirin can induce oxidative stress effects in GES-1cells. Rebamipide can activate Nrf2/HO-1pathway and relieve aspirin-induced oxidative stress in GES-1cells.