Studies On Antioxidant Activity Of Oligosaccharide Complexes In ARPE-19 Cells And Mice

Retinal pigment epithelial?RPE?cell degeneration is a clinical marker of age-related macular degeneration?AMD?.With the mitochondrial oxidative phosphorylation reduction,RPE cell metabolism and dysfunction,leading to AMD development.The oxidative damage of RPE cells is the main cause of AMD pathology.In this paper,the oxidative damage model of ARPE-19 cells was established by H₂O₂.The activity of oligosaccharide complex?OSC?was detected by cell viability,morphological observation of apoptosis,SOD activity,GSH and MDA content,ROS and mitochondrial membrane potential detection in ARPE-19 cells.The results showed that the oxidative stress of ARPE-19 cells induced by H₂O₂ was attenuated in a dose-dependent manner after using different concentrations of OSC.When the OSC concentration was 250?g / mL,the cell viability increased 19%,and the apoptotic morphology was weakened.SOD activity and GSH content were recovered to 94.8%and 90.1% of the control group respectively.MDA content and ROS content decreased to 1.47 times and 1.4 times of the control group respectively,and the mitochondrial membrane potential was significantly increased.This study shows that OSC has a certain antioxidant capacity in ARPE-19 cells.Then,the oxidative damage model of mice was established by ethanol.The antioxidant activity of OSC on ethanol-injured mice was investigated by the activity of SOD,the contents of GSH and MDA in serum.The results showed that when the OSC was given 0.8g / kg,the activity of SOD and the content of GSH were restored to 94.1% and 100.1% of the control group respectively,while the MDA content decreased to 1.49 times of the control group.The results of this action show that OSC in mice also have a good antioxidant function,can withstand the ethanol caused by oxidative damage to mice.Finally,in order to study the antioxidant mechanism of OSC,we used Western Blot to detect the expression levels of SIRT1 and PGC-1? in ARPE-19 cells and the liver and eye of mice.The expression of SIRT1 and PGC-1? in ARPE-19 cellsincreased by 31.6% and 23.7% compared with the H₂O₂ group respectively,when OSC concentration was 250?g/m L.When the OSC was given 0.8g/kg,the expression of SIRT1 protein in mice liver and eye were increased by 23.3% and 35.2%,respectively,and the expression of PGC-1? was up to 13.6% and 11.5% respectively compared with the control group.Indicating that OSC can significantly activate the SIRT1 / PGC-1? signaling pathway to withstand oxidative damage.In summary,OSC has the function of protecting H₂O₂ oxidized injured ARPE-19 cells and ethanol-oxidized mice by activating the SIRT1 / PGC-1? signaling pathway against oxidative damage.Therefore,OSC can be a potential drug for the prevention and treatment of AMD.