The Protective Mechanism Of Astragaloside In Oxidative Stress-induced PC12Cell Injury

Objective Astragaloside IV (AstragalosideIV, AsIV) has a variety of biological activity,such as antioxidant, free radical scavenging,anti-inflammatory and other pharmacologicaleffects.The aim of this article is to demonstrate the mechanism of puerarin in oxidativestress induced PC12cells injury by molecular biological methods.Methods In this study,H2O2were used to induce PC12cells,mitochondrial damage.Cell activity was detected by MTT.Mitochondrial swelling was measured in order todetect the mPTP opening,which was evaluated by the decrease of absorbance(OD).Cellular signaling factors such as GSK-3β and VASP were tested by western blot.Results1MTT assay was used to detect cell activity. Compared to the H2O2group,The absorbance ratio of control group decreased markedly(53.2±3.7%,P<0.05).Compared to the H2O2group,cells treated with different concentrations of AsIV(100,200,400nmol/L) showed a less decrease in absorbance ratio(70.4±5.1%,P<0.05;66.5±6.0%,P<0.05;61.9±3.8%,P<0.05),suggesting that AsIV could prevent oxidativestress induced mitochondrial injury.In addition,100nmol/L was the most effectiveconcentration (70.4±5.1%, P<0.05).2Mitochondrial swelling experiment results showedthat compared with the control group, absorbance (OD) values of H2O2(300umol/L)group decrease significantly(0.01380±0.00264,P<0.05).Compared to the H2O2group,100nmol/L AsIV pretreatment can inhibit H2O2resulted in reduction in ODvalues(0.01070±0.00250,P<0.05),suggesting AsIV may play a protective role byinhibiting mPTP opening.3Compared to the H2O2group,AsIV(100nmol/L) significantlyincreased the phosphorylation of GSK-3β(131.4±8.3%,P<0.05),and PKG inhibitorKT5823blocked the role of AsIV on GSK-3β phosphorylation(60.5±6.6%,P<0.05).Further Western blot results showed that compared with control group,AsIVsignificance increased the phosphorylation of VASP(Ser239)(116.0±7.1%, P<0.05).Thatwas abolished by KT5823(85.2±7.5%, P<0.05),further supporting that the cGMP-PKGsignaling pathway was responsible for inactivation of GSK-3β by AsIV.Conclusions1Astragaloside can reduce oxidative stress-induced neurotoxicity in PC12cells.2Astragaloside preven oxidative stress by inhibiting mPTP opening.3Astragalosideplay a protective role in PC12cells by the inactivation of GSK-3β and the inhibition of mPTP opening.4Astragaloside,which inhibit GSK-3β activity, may be related to cGMP/PKG signaling pathway.