Protective Effect of Shexiang(麝香)on PC12 Cells Oxidative Stress Injury Induced by Aβ1-42
Objective:To explore the protective effect of Shexiang(麝香)on oxidative stress injury and apoptosis in Alzheimer's disease(AD)cell models.Methods:The chemical components of Shexiang were analyzed by LC-MS/MS assay.An in vitro AD cell model was built by Aβ1-42 induced PC12 cells,and the survival rate of PC12 cells was determined by the CCK-8 assay to explore the cytotoxicity of Aβ1-42 on PC12 and the protective effect of Shexiang on Aβ1-42 induced PC12 cell injury.The effect of Shexiang on Aβ1-42 induced apoptosis in PC12 cells was de-tected by Annexin V-FITC/PI double staining,and that of Shexiang on Aβ1-42 induced mitochondrial membrane potential in PC12 cells was de-tected by the JC-1 method.The effect of Shexiang on reactive oxygen species(ROS)in Aβ1-42 induced PC 12 cells was detected by the DCFH-DA probe.Superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)kits were utilized to determine the effect of Shexiang on the activity levels of antioxidant enzyme in Aβ1-42 induced PC12 cells.Western blot was applied to detect the expressions of relevant apop-totic proteins Bax,Bcl-2,Cleaved caspase-3,Nuclear factor erythroid 2-related factor 2(Nrf-2),Heme oxygenase 1(HO-1),and Kelch-like ECH-associated protein 1(Keap 1)in PC 12 cells.Results:The seven chemical components of Shexiang were identified by LC-MS/MS:Hy-poxanthine,Androstenedione,Epitestosterone,Hexadecanamide,Muscone,Oleamide,and Stearamide.Compared with the blank control group,the model control group achieved a significantly reduced survival rate of PC12 cells(P＜0.01),significantly increased intracellular ROS level and apoptosis rate(P＜0.01),significantly decreased mitochondrial membrane potential(P＜0.05),and significantly reduced activities of SOD and GSH-Px(P＜0.05 or P＜0.01).The expressions of Bax,Cleaved caspase-3,and Keap 1 protein were significantly upregulated,while the expressions of Bel-2,Nrf-2,and HO-1 protein were significantly downregulated(P＜0.05 or P＜0.01).Compared with the model control group,the survival rate of cells was significantly increased in the 0.025,0.05,and 0.1 mg/mL Shexiang groups(P＜0.05 or P＜0.01),the intracellular ROS level and apoptosis was significantly reduced(P＜0.01),and the activity of SOD was significantly increased(P＜0.05 or P＜0.01).Specifically,mitochondrial membrane potential was significantly increased(P＜0.01)and the protein expressions of Bax,Cleaved caspase-3,and Keap 1 were significantly downregulated(P＜0.05 or P＜0.01)in the 0.05 and 0.1 mg/mL Shexiang group.In addition,the activity of GSH-Px was significantly increased(P＜0.01)in the 0.1 mg/mL Shexiang group,and protein expressions of Bcl-2,Nrf-2,and HO-1 were significantly upregulated(P＜0.05 or P＜0.01).Conclusion:Shexiang significantly improves the oxidative stress and apoptosis of the Aβ1-42 induced AD cell model through the Nrf-2/HO-1 pathway.
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