Puerarin Inhibits The Effects Of Ferroptosis On Neurological Impairment In Parkinson’s Disease

Objective:Parkinson’s disease(PD)is a neurodegenerative disease characterized by the loss of dopaminergic neurons,and its pathogenesis is still unclear.Ferroptosis is an iron-dependent form of cell death,and studies have suggested that ferroptosis is related to the pathogenesis of PD.This study aims to demonstrate that Puerarin inhibits ferroptosis and alleviates neurological impairment in PD.Methods:Firstly,the mouse model of PD was established by intraperitoneal injection of MPTP,and then the experiment was divided into the following five groups:the control group,MPTP group,12.5 mg/kg Pue+MPTP group,25 mg/kg Pue+MPTP group and 25 mg/kg Pue+MPTP group.The changes of motor impairment of PD mice were evaluated by Pole test,Traction test and Open field test.HE,Nissl and immunohistochemical staining were used to observe the pathological changes of brain tissue in PD mice.The content of dopamine(DA)in striatum was analyzed by HPLC.In addition,SH-SY5Y cells were induced by 20μmol/L Erastin to construct ferroptosis models in vitro,and they were divided into the following five groups according to the experimental protocol:the control group,Erastin group,20μmol/L Pue+Erastin group,40μmol/L Pue+Erastin group and 60μmol/L Pue+Erastin group,and then the survival rate of SH-SY5Y cells was detected by CCK-8 method.The content of intracellular iron,glutathione(GSH),reactive oxygen species(ROS)and lipid Peroxidation(LPO)were detected by the kits respectively,and then Western blot was used to detect the expression of glutathione peroxidase 4(GPX4)and xCT protein in each group.SH-SY5Y cells were induced by 3mmol/L MPP~+to build a model of PD nerve cell damage in vitro,and then experimental groups and Pue concentrations were consistent with the cell model induced by Erastin.The cell survival rate,iron,GSH and MDA contents of each group were measured respectively,and the structural changes of mitochondria in cells were observed by transmission electron microscope(TEM),and finally,the expressions of ferroptosis-related proteins xCT/GPX4 and iron homeostasis proteins in the substantia nigra of PD mice were detected by Western blot.Results:The results of experiments in vivo showed that Pue could improve the dyskinesia of PD mice,reduce the loss of dopaminergic neurons in substantia nigra,and reduce the damage of nerve function to a certain extent.The results of in vitro experiments showed that Pue could regulate the xCT/GPX4 pathway to reduce the levels of GSH,iron and LPO,and inhibit Erastin-induced ferroptosis.In addition,in vitro and in vivo models of PD have also shown that Pue can inhibit the increase of ferroptosis in MPP~+-induced cells to a certain extent,and up-regulate the expressions of xCT and GPX4 in the substantia nigra of PD mice.The expression of iron homeostasis proteins TfR1,FPN1 and FTH1 reduces iron deposition in the substantia nigra and inhibits neuronal ferroptosis.Conclusion:The data of this study preliminarily confirmed that Pue may regulate the xCT/GPX4 pathway and iron homeostasis to inhibit neuronal ferroptosis and reduce neurological damage in Parkinson’s disease.