Sulforapahne Protects Neurons Against Injury Caused By Oxygen-glucose Deprivation/reoxygenationand Its Mechanism

Objective:To investigate whether SFN protects neurons against injury caused byOxygen-Glucose Deprivation (OGD)/Reoxygenation(R) and the possiblemechanisms involved in neuroprotection.Methods:(1) Primary neuronal cultures of cerebral cortex were obtained from1-day-old Sprague-Dawley rats on day5-6in vitro.(2) The cortical neurons were exposed to OGD for1h，followed byreoxygenation for24h and were treated with0.1,0.2,0.5,1,2.5,5μmol/LSFN for25h during OGD/R. A PI3K specific inhibitor(LY294002)(10μmol/L) was added to cortical neurons with or without SFNfor25h during OGD/R. The cells were randomly divided into control group,OGD group, SFN group, SFN+LY group and LY group.(3)After24h reoxygenation, MTT was used to quantify cell viabilityassay; cell injury was measured by Hoechst33258/propidium iodide(PI) staining; immunofluorescence staining and western blot were used to detectexpression on molecular events related to apoptosis.Results:(1) MTT assay showed that1μmol/L SFN significantly increased cellviability;(2) While, Hoechst33258/PI staining showed that injured neurons wasreduced significantly in the SFN group.(3) Furthermore, immunofluorescence staining and western blotshowed that SFN increased Bcl-2expression and decreased cleavedcaspase-3expression. LY294002inhibited phosphorylated-Akt (p-Akt)expression evoked by SFN, decreased Bcl-2expression，increased cleavedcaspase-3expression.Conclusion:Together, these results suggest that SFN protects neurons againstinjury caused by OGD/R and this neuroprotective effect may be partlyassociated with PI3K/AKT anti-apoptotic pathway.