The Mechanism Of Asiaticoside On Inhibiting Oxidative Stress Induced By High-glucose Peritoneal Dialysis Solution In Human Peritoneal Mesothelial Cells

OBJECTIVE:To observe the effects of Asiaticoside(AS)on oxidative stress and activation of p38 mitogen-activated protein kinase(p38MAPK)pathway in human peritoneal mesothelial cells induced by high-glucose peritoneal dialysis solution(PDS),and to explore the molecular mechanism of Asiaticoside on peritoneal antioxidant stress.METHORDS:The human peritoneal mesothelial cell line HMrSV5 cells were cultured in vitro.We used CCK-8 method to detect the effects of 4.25%glucose peritoneal dialysis solution and different concentrations(10、50、100、150、200μmol/L)of AS on cell viability.The appropriate concentrations of AS for the follow-up experiments were filtered according to the results of CCK-8 detection on cell viability.Then the cells were randomly divided into 7 groups:control group,PDS group,AS low(50μmol/L),medium(100μmol/L)and high(150μmol/L)dose group,NAC group and SB203580 group.The cells were respectively cultured for 6,12 and 24 hours.The cells were stained with 2 ’,7 ’-Dichlorofluoresic diacetate(DCFH-DA)and the production of reactive oxygen species(ROS)were detected by flow cytometry.The level of malondialdehyde(MDA)and total-superoxide dismutase(T-SOD)in cells were measured by colorimetric method.Western blot was used to detect the expression of phosphorylated p38MAPK(p-p38MAPK)and p38MAPK protein.RESULTS:①Under the effects of PDS,the cell viability of HMrSV5 decreased,the ROS production increased and the p-p38MAPK expression was up-regulated.All these results were most obvious with the stimulation of PDS for 24 hours.The level of MDA increased while the level of T-SOD decreased in cells with the stimulation of PDS for 24 hours;② AS low,medium and high dose group could improve the effect of PDS on cell viability and reduce ROS generation.AS medium and high dose group could decrease the level of MDA,increase the level of T-SOD and down-regulate the p-p38MAPK expression under the stimulation of PDS.All these effects of AS were dose-related and the most significant effects were created in the AS high dose group;③The antioxidant NAC and p38MAPK inhibitor SB203580 could improve the effect of PDS on cell viability,reduce the production of ROS,decrease the level of MDA,increase the level of T-SOD and down-regulate the p-p38MAPK expression.CONCLUSION:High-glucose peritoneal dialysis solution can activate p38MAPK signaling pathway and induce oxidative stress in human peritoneal mesothelial cells.Asiaticoside is likely to reduce the the level of oxidative stress induced by high-glucose peritoneal dialysis solution in human peritoneal mesothelial cells by inhibiting p38MAPK signaling pathway to play protective effect on the peritoneum.