Effects of scutellarin on PKCγ in PC12 cell injury induced by oxygen and glucose deprivation

Aim:To evaluate the neuroprotective effect and mechanisms of scutellarin (Scu)against PC12 cell injury after oxygen and glucose deprivation followed byreperfusion (OGD-Rep).Methods:Undifferentiated rat pheochromocytoma PC12cells,exposed to oxygen and glucose deprivation followed by reperfusion (OGD-Rep),used as an in vitro model of ischemia/reperfusion.Cell survival was evalu-ated by diphenyltetrazolium bromide (MTT) assay and the amount of LDH releasewas determined using assay kits.[Ca2+]iwas monitored using a fluorescent Ca2+-sensitive dye Fura-2 acetoxymethyl ester.Cell apoptosis was detected by aDNA ladder and by flow cytometric detection.The expression of protein kinaseC (PKC)γ was determined using both RT-PCR and Western blotting.The trans-location of PKCγ was assayed by subcellular fractionation and Western blotting.Results:OGD-Rep injury significantly elevated the level of LDH release,[Ca2+]i,mRNA expression and the translocation of PKCγ compared in the PC12 cellswith those of the normal group.Scu (10-100 μmol/L) exerted a protectiveeffect against OGD-Rep injury by reducing LDH release,[Ca2+]i,the percent ofapoptosis,and the translocation of PKCγ.Conclusion:Scu inhibits the increaseof [Ca2+]iand the activation of PKCγ,exerting protective effects against PC12cell injury induced by OGD-Rep.