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 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 evaluated by diphenyltetrazolium bromide （MTT） assay and the amount of LDH release was 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 a DNA ladder and by flow cytometric detection. The expression of protein kinase C （PKC）T was determined usiiag both RT-PCR and Western blotting. The translocation of PKCT 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 cells with those of the normal group. Scu （10-100 μmol/L） exerted a protective effect against OGD-Rep injury by reducing LDH release, [Ca^2＋]i, the percent of apoptosis, and the translocation of PKCγ. Conclusion： Scu inhibits the increase of [Ca^2＋]i and the activation of PKCγ, exerting protective effects against PC12 cell injury induced by OGD-Rep.

Effects of scutellarin on PKCgamma 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 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 evaluated by diphenyltetrazolium bromide (MTT) assay and the amount of LDH release was determined using assay kits. [Ca2+](i) was monitored using a fluorescent Ca2+-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)gamma was determined using both RT-PCR and Western blotting. The translocation of PKCgamma 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 PKCgamma compared in the PC12 cells with those of the normal group. Scu (10-100 micromol/L) exerted a protective effect against OGD-Rep injury by reducing LDH release, [Ca2+](i), the percent of apoptosis, and the translocation of PKCgamma. CONCLUSION: Scu inhibits the increase of [Ca2+](i) and the activation of PKCgamma, exerting protective effects against PC12 cell injury induced by OGD-Rep.