The Protective Effect Of F-NBP, A Derivative Of Dl-3-n-Butylphthalide, On Hydrogen Peroxide-induced Oxidative Damage In PC12 Cells

Objective:To investigate the protective effect of 3-butyl-6-fluoro-1(3H)-isobenzofuranone on the H2O2-induced cytotoxicity in rat pheochromocytoma cells (PC12) cells.Methods:Different dose of Na2S2O4 was used to induce the cytotoxicity in PC 12 cells in order to optimize dose by MTT assay. Different dose of H2O2 was used to induce the cytotoxicity in PC 12 cells in order to optimize dose by MTT assay. The enzyme activity of superoxide dismutase(SOD), glutathione peroxidase(GSH-PX) and the lipid peroxidation products malondialdehyde(MDA) in the cells were observed. DNA-binding dye Acridine Orange(AO) was used to determine the morphological characteristic of apoptotic cells. PI was used to a signature of cell membrane integrity and cell damage.Mitochondrial membrane potential in PC 12 cells were monitored by fluorospectrophotometer combining with Rh123.Results:1. After treatment with 2mM Na2S2O4 in PC 12 cells for 24 hours, the cell viability decreased markedly, while F-NBP at the different concentration (5μm,10μM,50μM) could elevate the cell viability obviously.2. After treatment with 500μM H2O2 in PC 12 cells for 24 hours, the cell viability decreased markedly, while F-NBP at the different concentration (5μM,10μM,50μM) could elevate the cell viability obviously.3. The results suggest that H2O2 decreasedGSH-PX and GSH-PX activity, in contrast, H2O2 increased MDA and ROS. F-NBP significantly decreased ROS and MDA, increased GSH-PX activity in PC 12 cells.4. After the treatment with 500μM H2O2, the mitochondrial membrane potential significantly decreased, microscopic observation showed that PC 12 cells exhibited morphological alterations such as cell shrinkage and membrane blabbing, the typical characteristics of apoptotic cell death, which was reduced with co-treatment of F-NBP.Conclusions:1.2 mM Na2S2O4 could induce ischemia and apoptosis in PC 12 cells, which result in the cell death ultimately.2.500μM H2O2 could induce toxicity, oxidative damage and apoptosis in PC 12 cells, which result in the cell death ultimately.3. The study shows that F-NBP at the different concentration has a protective effects against H2O2-induced cytotoxicity and apoptosis in PC 12 cells, which may represent the cellular mechanisms including scavenging ROS, increasing GSH-PX activity. F-NBP may also represent a potential treatment strategy for ischemia.