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

<正> 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+]i was 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+]i and the activation of PKCγ,exerting protective effects against PC12cell injury induced by OGD-Rep.