Effect Of Anacyclus Pyrethrum Extract On Hydrogen Peroxide Induced Injury In PC12 Cells

Objective: To study antioxidant effect and related mechanism of traditional Uyghur medicine drug Anacyclus pyrethrum extracts on Hydrogen peroxide-induced apoptosis in PC12 cells and elucidate the mechanisms and thrapeutic effects of Anacyclus pyrethrum on Alzheimer's disease.Methods: Water and alcohol extracts of Anacyclus pyrethrum were extracted according to the extraction method.PC12 cells were cultured and selected its logarithmic growth phase by CCK-8 cell proliferation assay.CCK-8 method was used to detect effect of different concentrations of Hydrogen peroxide on cell proliferation in time dependent manner and also was used to build experimental model of cell injury induced by Hydrogen peroxide based on optimum injury time and concentration.Cultured cells were randomly divided into normal control group,model group,Anacyclus pyrethrum water extract group,Anacyclus pyrethrum alcohol extract group and positive control group to measure the cell viability by CCK-8 cell proliferation assay.The degree of Hydrogen peroxide induced PC12 cell apoptosis was characterized by using Hoechst 33342 staining and PE Annexin V staining.PC12 cells Apoptosis was evaluated by LDH,MDA,SOD release.Under the laser confocal microscope,the changes of mitochondrial membrane potential of each group were observed by Rhodamine.Expression of Bax,Caspase-3 and Bcl-2 protein in PC12 cells of each group was assessed by Western Blotting.Results: The results showed that Anacyclus pyrethrum extract promoted PC12 cell proliferation,inhibited enhanced apoptosis rate.Compared with model group the value of cell proliferation of Anacyclus pyrethrum extract groups with different concentration statistically had significant difference(P<0.05).Hochest 33342 staining found that cell morphology and number of cells that are treated with different concentrations of extract groups close to the normal group and differ from model group.Annexin-V/PE double staining showed that apoptosis rate of the model group was higher than normal group and the early apoptosis rate of the cells in different concentration extrat groups was less than model group(P<0.01).Results of enzyme assay showed that significantly reduced LDH,MDA level and increased SOD level,and protected PC12 cells from Hydrogen peroxide induced cytotoxicity.Mitochondrial membrane potential detection showed that compared with the normal group,the intracellular fluorescence of model group was significantly weakened.Contrary to model group,intracellular fluorescence of different concentrations of Anacyclus pyrethrum extract group was increased.Protein expression test showed that compared with the normal group,the expression of Bcl-2 protein in model group was decreased and the expression of Caspase-3 and Bax protein was enhanced.Compared with the model group,the expression of Bcl-2 protein in extract group was increased,and the expression of Bax and Caspase-3 protein significantly decreased.Conclusion: Anacyclus Pyrethrum extract clearly reduces percentage of apoptosis cells and protects Hydrogen peroxide induced cell injury by regulating oxidative stress in PC12 cells.