Expression Of ATP-P2X4 Signal Axis In PD Animal Model

Background:Parkinson’s disease is a common neurodegenerative disease in the elderly.It is characterized by the progressive loss of dopaminergic neurons.The pathogenesis of Parkinson’s disease is still unknown.Recent studies have shown that abnormal activation of microglia plays an important role in the pathogenesis of Parkinson’s disease mediated by neuroinflammatory mechanism.Microglia can be activated abnormally under adenosine triphosphate(ATP)stimulation,which mediates neuronal injury by secreting a large number of inflammatory factors(such as tumor necrosis factor-α,IL-1 β,etc.).Inhibiting the abnormal activation of microglia and effectively inhibiting the immune inflammatory injury may be an effective new approach in the treatment of Parkinson’s disease.P2X4 R,as a ATP receptor on microglia,may be a new target to inhibit abnormal activation of microglia and inhibit the development of Parkinson’s disease.The purpose of this study was to study the expression of ATP-P2X4 signal axis in PD animal model and its effect on the inflammatory factor TNF-a,and to provide a new target for the prevention and treatment of PD.Methods:All the Wistar rats were randomly divided into two groups: 6-OHDA group and Control group.6-OHDA group was injected with 6-OHDA into the left substantia nigra of rats by stereotactic locator and microinjector,and the control group was injected with saline containing 2% ascorbic acid into the left substantia nigra by stereotactic instrument and microinjector.After the establishment of the model,the behavior of rats was observed by intraperitoneal injection of APO to induce rotation,and the PD rats were selected successfully.After 4 weeks of modeling,the samples of substantia nigra and striatum were collected and perfused into the brain,then the number of dopaminergic neurons in the substantia nigra was determined by th immunofluorescence staining.The expression levels of P2X4 R,NLRP3,th,TNF-α,Caspase-1 and IL-1 β in substantia nigra were detected by Western blot method.The expression levels of P2X4 R,NLRP3,th,TNF-α,Caspase-1 and IL-1 β m RNA in substantia nigra were assessed by RT-PCR.Results:The number of dopaminergic neurons and the expression of th in substantia nigra in 6-OHDA group were significantly lower than those in Control group.The expression of P2X4 R,NLRP3,th,TNF-α,Caspase-1 and IL-1 β in 6-OHDA group was significantly higher than that in Control group.Conclusions:In this experiment,the expression of P2X4 R,NLRP3,TNF-α,Caspase-1,IL-1 β protein and th positive cells in substantia nigra of PD model rats prepared by6-OHDA was detected.It was proved that P2X4 has a role in the pathogenesis of PD.It was confirmed that the signal axis of ATP-P2X4 and its downstream NLRP3 inflammatory corpuscles,TNF-α,Caspase-1,IL-1 β inflammatory factors had significant changes,indicating that the signal axis may play an important role in the pathogenesis of Parkinson’s disease through the mechanism of neuroinflammation.Regulation of this signal axis provides a new possibility for the targeted therapy of PD,and also provides a requirement for further research.