Effect Of Overexpression Of P2X4 On Apoptotic Protease In Rat Brain Of Parkinson’s Disease Model

Background and objective: Parkinsonian disease(PD)is the second most common neurodegenerative disease in Alzheimer’s disease(AD).In previous studies,it is believed that environmental factors,genetic factors,aging,oxidative stress,mitochondrial function failure,calcium overload and other factors are the result of synergy.Recently,the role of neuro-immune inflammation in the pathogenesis of PD has been widely concerned by researchers.The study on the regulation mechanism of related signal pathway is of great significance to elucidate the pathogenesis of PD and can provide a direction for the prevention and treatment of PD.The activation of NLRP3 inflammatory corpuscles mediated by ATP-P2X4 R signaling axis is one of these key pathways.The role of neuroimmune inflammation mediated by ATP-P2X4 R signal axis in the pathogenesis of PD was clarified.Overexpression of P2X4 R gene may enhance the inflammatory response of PD by up-regulating the expression of signal pathway mediated by P2X4 R,and enhance the injury of DA neurons.The protective effect of inhibiting P2X4 R mediated inflammatory pathway on PD was proved from the side.To investigate the effect of overexpression of P2X4 gene on the expression of apoptotic protease caspase-1 in rat model of Parkinson’s disease induced by 6-OHDA.To reveal the role of P2X4 in the pathogenesis of PD and to provide a new target for the prevention and treatment of PD.Methods: one hundred and twenty male Wistar rats weighing about 200-250 g were randomly divided into 6 groups: the 6-OHDA group;the control group;the target gene RNA-P2X4+6-OHDA group;the target gene RNA-P2X4 group;the negative virus P2X4-NC+6-OHDA group;the pure negative virus P2X4-NCgroup.P2X4-NC lentivirus carrying empty RNA and up-regulation of P2X4 objective RNA-P2X4 lentivirus was injected into the left substantia nigra of rats via a stereoscopic locator.According to the group situation,6-OHDA-8ug-2ul)or saline(only 0.2% ascorbic acid 2ul)was injected into the left substantia nigra of rats by brain stereotactic locator.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,the rat head was cut off,and the changes of th in substantia nigra were detected by immunofluorescence staining,and the expression of P2X4 R,NLRP3 and caspase-1 protein were detected by Western blot.The expression levels of P2X4 R,NLRP3 and caspase-1 m RNA were detected by RT-PCR method.Results: compared with the P2X4-NC+6-OHDA group,the expression of P2X4R(1.099±0.05569 vs 0.7821±0.02008,P(27)0.001),NLRP3(0.9875±0.01932 vs 0.6645±0.01747,P(27)0.001),caspase-1(0.9948±0.01788 vs 0.8276±0.04543,P(27)0.001)were significantly increased in P2X4-NC+6-OHDA group.Conclusion: overexpression of P2X4 gene can increase the expression of caspase-1 protein in PD dopaminergic neuron model,promote apoptosis response,and have potential neuropathic effects.P2X4 plays an important role in the occurrence and development of PD.