6-O-Galloylpaeoniflorin Attenuates Cerebral Ischemia Reperfusion-Induced Neuroinflammation and Oxidative Stress via PI3K/Akt/Nrf2 Activation

Figure 2

GPF protected the hypoxic-ischemic injury in PC12 cells. To determine the neuroprotective effects of GPF, PC12 cells were pretreated with different doses of GPF (0, 10, 50, or 100 μM, resp.) for 1 h before being subjected to OGD. PC12 cells without GPF and OGD treatment were set as a control. The cellular injury induced by OGD was analyzed by a cellular viability assay (CCK-8 assay) (a) and a LDH activity assay (b). GPF treatment at the indicated doses relieved OGD-induced cellular injury. The amount of apoptotic PC12 cells was determined by TUNEL staining. TUNEL-positive and total (DAPI-positive) number of the cells in each group were counted manually by two independent observers in 6 random microscopic fields (400x), and the percentage of TUNEL-positive cells were subsequently calculated (c). GPF decreased the ratio of TUNEL-positive cells (purple fluorescence, labeled with white arrows) in a dose-dependent manner. Nuclei were stained by DAPI (blue fluorescence). GPF downregulated the OGD-induced levels of caspase-3 in a dose-dependent manner in PC12 cells. The data of each group were from three independent assays. versus 0 mg/kg GFP with OGD; versus 0 mg/kg GFP with OGD.

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