The Protective Effects And Possible Mechanisms Of 6-gingerol Against Hydrogen Peroxide Induced DNA Damage In INS-1 Cells

Objective:With the change of life style and dietary constituents,the number of patients with diabetes was increasing year by year.Diabetes has become one of the largest global health emergencies of the 21 st century.6-Gingerol,one of the most important activity components in gingers,has a wide range of pharmacological effects,such as anti-oxidation,anti-inflammatory,anti-tumor,regulating blood sugar and so on.6-Gingerol has broad application prospects in the prevention and treatment of diabetes and its complications,but its specific mechanism is not very clear.The present study was designed to investigate the molecular mechanism of 6-gingerol in the prevention and treatment of diabetes mellitus at the cellular level,through observed the protective effects of 6-gingerol against hydrogen peroxide induced toxicity in INS-1 cells.It would provide the necessary basis for the development of new therapies for diabetes mellitus and the development of new medicines and health foods.Method:INS-1 cells were selected as the research objects in this study.The INS-1 cells were treated with H₂O₂?100 ?M?for 1 h,resulting in oxidative damage of cells.6-Gingerol was used as a protective agent.The INS-1 cells were pretreated with 6-gingerol for 24 h before treated with H₂O₂,and then we observe the protective effects of different doses of 6-gingerol?2.5 ?M?5 ?M?10 ?M?on cell injury.Single Cell Gel Electrophoresis?SCGE?assay was used to detect DNA strand breaks in INS-1 cells;the levels of intracellular ROS and GSH,the stability of lysosomal membrane and mitochondrial membrane potential were measured by fluorescence spectrophotometer;RT-PCR test was used to detect the expression levels of ATM and p53 genes.Results:Compared with the control group,the INS-1 cells showed comet-like tails after treated with H₂O₂?100 ?M?1 h,and the tail length,tail DNA / head DNA?%?and tail moment were significantly increased?P < 0.01?.Pretreated with 6-gingerol for 24 h before treated with H₂O₂?100 ?M?1 h,the comet-like tails of INS-1 cells were reduced,that tail DNA / head DNA?%?decreased significantly?P < 0.05 or P < 0.01?and the tail length and tail moment decreased significantly?P < 0.01?.Compared with the control group,after treated with H₂O₂?100 ?M?1 h in the INS-1 cells,the level of intracellular ROS were increased?P < 0.01?and the levels of GSH were decreased?P < 0.01?.The cells were in the state of oxidative stress.Compared with the H₂O₂ group,after pretreated with different doses of 6-gingerol for 24 h before treated with H₂O₂?100 ?M?1 h,the levels of intracellular ROS were decreased significantly?P < 0.05 or P < 0.01?and the levels of GSH were increased significantly?P < 0.05 or P < 0.01?.Compared with the control group,after treated with H₂O₂?100 ?M?1 h in the INS-1 cells,the stability of lysosomal membrane were decreased significantly?P < 0.01?and mitochondrial membrane potential were decreased significantly?P < 0.01?.Compared with the H₂O₂ group,after pretreated with different doses of 6-gingerol for 24 h before treated with H₂O₂?100 ?M?1 h,the stability of lysosomal membrane were increased significantly?P < 0.01?and mitochondrial membrane potential were increased significantly?P < 0.05?.Compared with the control group,after treated with H₂O₂?100 ?M?1 h in the INS-1 cells,the expressions of ATM and p53 in cells were increased significantly?P < 0.01?.Compared with the H₂O₂ group,after pretreated with different doses of 6-gingerol for 24 h before treated with H₂O₂?100 ?M?1 h,the expressions of ATM and p53 in cells were decreased significantly in a dose-dependent manner?P < 0.05 or P < 0.01?.Conclusion:1?H₂O₂ could induce DNA damage in INS-1 cells;as an antioxidant,6-gingerol could protect the damage of INS-1 cells.2?6-Gingerol may play an important role in the protection of INS-1 cells by the lysosomal-mitochondrial pathway and acting on the ATM-p53 dependent signaling pathway.