| Parkinson’s disease is second only to alzheimer’s disease of the world’s second largest central neurodegenerative diseases,numerous studies have proved to be the substantia nigra in the brain striatum lesions occur,its main cause is not yet clear,general with multiple factors such as heredity,environment and aging,through the defect in the proteasome dysfunction,mitochondrial function,oxidative stress,inflammatory reaction makes the dopaminergic neuron death and the neurons to regenerate,eventually necrosis lose too many neurons,making inadequate secretion of dopamine,and disease.In recent years,with the emergence of the aging population,the incidence of PD in China has been on the rise year by year and there are a large number of young patients,which brings a heavy burden on the healthy life of patients and their families.At present,the means and drugs to treat PD can only slow down the effect but cannot cure it.Some common western drugs can maintain the level of dopamine in the brain of patients,but the side effects are quite large and drug resistance is easy to occur.In traditional Chinese medicine in the treasure house,there are a lot of drugs to treat shock syndrome type of diseases such as Parkinson’s disease and remarkable effect,but many prescription drugs on,such as single herbs because of complex chemical composition,interaction is unknown,a given medicine medicinal materials quality problem of the difference of homogeneity and failed to make in-depth research,to evaluate the effect.Therefore,this study only used the traditional Chinese medicine Acanthopanax acanthopanax to verify the effect and mechanism of Eleutheroside E extracted from Acanthopanax acanthopanax on PD oxidative stress cell model and inflammatory cell model.To provide theoretical and experimental basis for the clinical application of Eleutheroside E in PD,this paper includes the following main contents:(1)The effect and mechanism of Eleutheroside E on PD oxidative stress cell model: MPTP(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine)was used as inducer,and 2500μm MPTP was added into PC12 cells(rat adrenal pheochromocytoma cells)for 24 h to complete the model establishment.MTT and CCK-8 cell viability were used to determine the tolerance of PC12 cells to Eleutheroside E and the effect of Eleutheroside E on the cell viability of PD oxidative stress model.Mitochondrial membrane potential detection was used to determine the protection of Eleutheroside E against the mitochondrial membrane potential of PD oxidative stress model.Reactive oxygen ROS detection was used to determine whether Eleutheroside E reduced ROS content in the oxidative stress model of PD.By flow cytometry,Annexin V APC/PI double staining was used to determine the reduction of apoptosis rate of Eleutheroside E in the oxidative stress model of PD.Finally,the expression changes of Cyt C,Nrf2 and NQO1 of Eleutheroside E after addition were detected by Western blot to clarify the mechanism of Eleutheroside E on the oxidative stress model of PD.(2)Effect of Eleutheroside E on PD inflammatory response cell model: To establish the PD inflammatory response model,LPS(lipopolysaccharide)was used as inducer,and 1.25mg/ m L LPS was added to PC12 cells for 24 h to complete the model establishment.MTT and CCK-8 cell viability were determined to determine the effect of Eleutheroside E on the cell viability of the inflammatory response model of PD.Mitochondrial membrane potential detection was used to investigate whether LPS had an effect on the mitochondrial membrane potential of PC12 cells.By flow cytometry,Annexin V APC/PI double staining was used to determine the reduction of apoptosis rate of Eleutheroside E in the inflammatory response model of PD.Finally,ELISA immunoenzyme linked adsorption method was used to detect the changes in the contents of TNF-α and IL-6,two inflammatory factors secreted by the PD inflammatory response model after addition of drugs,to indicate the mechanism of action. |
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