The Protective Effect Of Velvet Antler Polypeptide On The Damage Of SH-SY5Y Cells Induced By Rotenone And Its Mechanism

Background:Parkinson’s disease(PD)is caused by the interaction of genetic factors and environmental toxins causing damage to dopamine(DA)energy neurons in the substantia nigra compact.The cytoplasm contains α-synuclein(α-synuclein).Synuclein(α-syn)Lewy body(lewy body),which in turn leads to abnormal degeneration and loss of DA neurons,a progressive neurodegenerative disease of DA levels in the brain.The pathogenesis of PD is complex and not yet fully clear.Current research indicates that the pathogenesis of PD involves mitochondrial dysfunction,oxidative stress,protein homeostasis,endoplasmic reticulum stress,autophagy and other mechanisms.The clinical symptoms of PD are mainly caused by the decrease of DA levels in the brain,so clinically mainly to improve the DA level of patients’ brains as the main treatment method.So far,oral levodopa is still the choice of most patients,but this will cause DA to The spatial and temporal distribution of the patient’s body is not regulated by the body,which easily causes side effects and is not conducive to the patient’s life and subsequent treatment.Therefore,it is necessary to seek new treatments.After thousands of years of accumulation and precipitation in China,Chinese medicine has a considerable understanding of common diseases.Its overall adjustment and syndrome differentiation also meet the current treatment needs of PD.There is a great deal of search for treatment methods in traditional medicine with the help of Chinese medicine theory The advantages.The clinical manifestations of PD are mainly static tremor,unresponsiveness,stiffness and twisted posture,etc.,which is very consistent with the traditional Chinese medicine tremor syndrome,and clinical Chinese medicine also treats PD as a tremor syndrome.Chinese medicine believes that the pathogenesis of PD is mainly liver and kidney deficiency,and treatment is also to nourish liver and kidney,regulate qi and blood,and mainly reduce phlegm and spasm.Deer antler is a traditional Chinese medicinal material that has the effect of invigorating the kidney and strengthening the bones.Modern research has found that velvet antler polypeptide has the ability to enhance superoxide dismutase activity and improve the efficiency of ROS clearance in the body,and can also promote the growth,proliferation and differentiation of neural stem cells.The purpose of this study was to use rotenone to induce SH-SY5Y cells to simulate DA neurons in PD patients,and to observe the effect and mechanism of antler polypeptide on SH-SY5Y cell injury model.Objective:To investigate the effect of antler polypeptide on rotenone-induced SH-SY5Y cell damage and related mechanisms.Methods:Use 0.5 μM rotenone to act on SH-SY5Y cells to establish an in vitro PD model.Set a blank group,model group,antler polypeptide high,medium and low dose group and positive control group.After the cells of each group were subjected to rotenone modeling and antler polypeptide intervention,the cell activity of each group was detected by MTT method to observe the effect of antler polypeptide on the cell proliferation activity of PD model;HE staining method was used to observe the effect of antler polypeptide on the morphology of PD model cell;Rhodamine 123(Rhodamine 123,Rh123)staining method was used to detect the mitochondrial membrane potential of cells,and the effect of velvet antler polypeptide on PD model mitochondrial dysfunction was observed.The effect of peptides on oxidative stress in PD model;immunohistochemical staining was used to detect the expression of α-syn,Akt and mTOR protein in cells,and the effect of velvet polypeptide on Akt/mTOR signaling pathway and α-syn protein in PD model influences.Results:HE staining results showed that compared with the blank group,the number of cells in the model group decreased,the antenna structure became dull,the shape became round,and eosinophilic Lewy bodies were visible in the cytoplasm.Compared with the model group,there was no significant difference in cell morphology between the positive control group and the antler polypeptide high,medium and low dose groups,but there were fewer Lewy bodies in the cytoplasm;Rhodamine 123 staining showed that compared with the blank group,The mitochondrial membrane potential of the model group decreased significantly(P<0.05).Compared with the model group,the mitochondrial membrane potential of the positive control group and the antler polypeptide high,medium and low dose groups were increased(P<0.05);DCFH-DA staining results showed that compared with the blank group,the model group cells The content of medium reactive oxygen species(ROS)increased significantly(P<0.05).Compared with the model group,the content of reactive oxygen species(ROS)in the positive control group and the antler polypeptide high,medium and low dose groups all decreased(P<0.05);immunohistochemical staining showed that compared with the blank group,the model group The expression of α-syn increased significantly(P<0.05).Compared with the model group,the expression of α-syn in the positive control group and the antler polypeptide high and medium dose groups was significantly reduced(P<0.05).Compared with the blank group,the expression of Akt in the model group was significantly increased(P<0.05).Compared with the model group,the expression of Akt in the positive control group and the antler polypeptide high and medium dose groups was significantly reduced(P<0.05).Compared with the blank group,the expression of mTOR in the model group was significantly increased(P<0.05).Compared with the model group,the expression of mTOR in the positive control group and the high-dose antler polypeptide group was significantly reduced(P<0.05)Conclusion:Antler polypeptide may promote the degradation of α-syn by inhibiting the Akt/mTOR signaling pathway,thereby exerting neuronal protection.