Study on the protective mechanism from Lycium ruthenium Murray anthocyanin monomer PrG against H2O2 induced oxidative damage in the granulosa cells of mice
Objdctive To investigate the protective mechanism of PrG,an anthocyanin monomer of Lycium ru-thenium Murray,against H2O2-induced oxidative damage in the granulosa cells(GCs)of mice.Methods The GCs from 20 female ICR mice were subjected to primary culture.Then,these mice were randomly divided into control group,hydrogen peroxide-treated group(H2O2group)and PrG-treated group(H2O2+PrG group).Firstly,an oxida-tive stress model of GCs induced by different concentrations(0 μmol/mL,25 μmol/mL,50 μmol/mL,100 μmol/mL,200 μmol/mL,400 μmol/mL)and different action times(30 min,60 min,120 min,180 min)of H2O2 was constructed.Subsequently,different concentrations of PrG(20 μg/mL,40 μg/mL,80 μg/mL)and different action times(12 h,24 h,48 h)were applied to treat GCs after oxidative damage.The optimal concentration and action time of H2O2 and PrG were selected by the CCK8 method.The proliferation of GCs induced by H2O2 was detected by the EdU method.Reactive oxygen fluorescence probe was used to detect reactive oxygen species(ROS)levels in GCs.Mmitochondri-al membrane potential and mitochondrial distribution were detected by using the JC-1 probe and Mito-tracker Red CMXRos live cell dye.Results The survival rate of mice GCs was decreased by about 60%at a concentration of 200 μM and a time of 2 h.The viability of GCs was significantly increased with the rising of PrG concentrations(P<0.01),and the optimal effect on the viability was observed at a concentration of 80 μg/mL.PrG significantly promoted the proliferative capacity of GCs,decreased the level of ROS(P<0.01),increased the number of mitochondria distribut-ed(P<0.01)and restored mitochondrial membrane potential(P<0.01).Conclusion The anthocyanin monomer PrG of Lycium ruthenium Murray exerted a protective effect on H2O2-induced oxidative damage of mouse GCs by scav-enging ROS free radicals and enhancing mitochondrial function.
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