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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.[Ca~(2+)]_i was monitored using a fluorescent Ca~(2+)-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,[Ca~(2+)]_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,[Ca~(2+)]_i,the percent ofapoptosis,and the translocation of PKCγ.Conclusion:Scu inhibits the increaseof [Ca~(2+)]_i and the activation of PKCγ,exerting protective effects against PC12cell injury induced by OGD-Rep.
Aim:To evaluate the neuroprotective effect and mechanisms of scutellarin (Scu)against PC12 cell injury after oxygen and glucose deprivation followed by reperfusion (OGD-Rep).Methods:Undifferentiated rat pheochromocytoma PC12 cells,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.[Ca~(2+)]_i was was used using A fluorescent Ca 2+ -sensitive dye Fura-2 acetoxymethyl ester. Cell apoptosis was detected by a DNA ladder and by flow cytometric detection. The expression of protein kinase C (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, [Ca 2+ ]]_i, mRNA expression and the translocation of PKCγ compared in the PC12 Cell Swith those of the normal group.Scu (10-100 μmol/L) exerted a protectiveeffect against OGD-Rep injury by reducing LDH release,[Ca 2+ ]_i, the percent of apoptosis, the translocation of PKCγ.Conclusion :Scu inhibits the increase of [Ca~(2+)]_i and the activation of PKCγ, exerting protective effects against PC12cell injury induced by OGD-Rep.