OBJECTIVE To investigate the protective effect proanthocyanidin B2(PC-B2)on oxidative damage and apoptosis of mouse astrocytes(AS)induced by hydrogen peroxide(H2O2)and its mechanism.METHODS AS were isolated and cultured from neonatal C57BL/6 mice(1-3 d).The optimal concentration of H2O2 and PC-B2 was divided into four groups:normal group,normal+PC-B2 group(100 μg·mL-1 PC-B2 treated for 24 h),H2O2 model group(200 μmol·L-1 H2O2 treated for 24 h),PC-B2 group(200 μmol·L-1 H2O2 and 100 μg·mL-1 PC-B2 treated for 24 h).The cell viability of each group was detected by CCK-8 method.Cytotoxicity was detected by LDH method.The antioxidant capacity was detected by ABTS and DPPH.The content of MDA and the activity of SOD,CAT and GSH-Px were detected by ELISA kit.Detection of apoptosis in each group was done by TUNEL staining.The mRNA and protein expression levels of Bax,Bcl-2,Caspase-3,Akt/Stat3,p-Akt,p-Stat3 and Nrf2/HO-1 in AS were detected by RT-PCR and Western blotting,respectively.RESULTS PC-B2 could significantly enhance cell viability and inhibit AS apoptosis.Compared with the H2O2 model group,PC-B2 intervention could significantly reduce the content of LDH and MDA in AS,and increase the activity of SOD,CAT and GSH-Px.PC-B2 intervention could inhibit the mRNA and protein expression of Bax and Caspase-3,and up-regulate the mRNA and protein expression of Akt/Stat3,Bcl-2,Nrf2/HO-1.CONCLUSION PC-B2 can enhance the antioxidant capacity of AS through Akt/Stat3 and Nrf2/HO-1 pathways,therefore reduce H2O2-induced AS oxidative damage and apoptosis.
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