The Effect Of ATP-P2X4 Signal Transduction On The Expression Of Il-1β And BDNF In Brachial Of Animals With Parkinson’s Disese

Objective The purpose of this study was to investigate the effects of lentivirus overexpression of P2X4 gene（LV-P2rx4）on the number of dopaminergic neurons and the protein relative expression levels of IL-1βand BDNF in substantia nigra of rats with Parkinson’s disease induced by 6-OHDA.In order to study the role of ATP-P2X4 signal axis and inflammatory factor IL-1βand neuroprotective factor BDNF in the pathogenesis of Parkinson’s disease.Methods The male Wistar rats with a body mass of 200～250 grams were randomly divided into 6 groups,with 20 rats each group.Stereotactic left midbrainsubstantia nigra.The control group was injected with saline containing 0.02%VitC.The6-OHDA group was injected with 6-OHDA.The RNA-P2X4 group was injected with lentivirus carrying overexpressed P2X4 gene.The P2X4-NC group was injected with airborne virus.The RNA-P2X4+6-OHDA group was injected with lentivirus carrying overexpressed P2X4 gene first,and then 6-OHDA a week later.The P2X4-NC+6-OHDA group was injected with airborne virus first,and then 6-OHDA a week later.14 days after the completion of the modeling,the behavioral test was performed by rotation testinduced by intraperitoneal injection of apomorphine.The average rotational speed≥7r/min was taken as the standard of modeling success.The successful model was selected into the experiment.After the model was successfully determined,the rats in each group were executed and the left substantia nigra tissues was collected quickly.The changes of the number of the TH positive neuron in the left substantia nigra of rats in each group was detected by immunofluorescence staining,Meanwhile,the protein relative expressionlevels of P2X4R,IL-1βand BDNF in the left substantia nigra of rats in each group were detected by Western blotting.Results The results of immunofluorescence staining detection show that:（1）Compared to the control group,the number of the TH positive dopaminergic neurons in the 6-OHDA group and P2X4-NC+6-OHDA group was reduced and the difference was statistically significant（P<0.01,F=3.79）.（2）Compared to the 6-OHDA group and P2X4-NC+6-OHDA group,the number of the TH positive dopaminergic neurons in RNA-P2X4+6-OHDA group was notably decreased,the difference was statistically significant（P<0.001,F=8.47）.The results of Western blot detection show that:（1）Compared to the control group,the protein relative expression of P2X4R and IL-1βincreased in the 6-OHDA group and P2X4-NC+6-OHDA group,while the relative expression of BDNF protein reduced and the difference was statistically significant(PP2X4R<0.01,FP2X4R=5.55;PIL-1_β<0.001,FIL-1_β=6.77;P_BDNF<0.05,F_BDNF=9.75).（2）Compared to the 6-OHDA group and P2X4-NC+6-OHDA group,the protein relative expression of P2X4R and IL-1βincreased in RNA-P2X4+6-OHDA group,but the BDNF protein decreased and the difference was statistically significant(P_P2X4R<0.001,F_P2X4R=8.47;P_IL-1β<0.001,_FIL-1β=13.75;P_BDNF<0.05,FBDNF=5.16).Conclusions Through our study,lentivirus-mediated P2X4 gene overexpression can reduce the number of the TH positive dopaminergic never cells,enhance the protein expression of IL-1βand decrease the expression of neuroprotective factor BDNF in the substantia nigra of rat brain.It is suggested that ATP-P2X4 signal axis can regulate the expression of IL-1βand BDNF.UP-regulate P2X4 gene expression can enhance the inflammatory response of microglia in substantia nigra of PD model rats,weaken the protective effect of neurokines,and damage dopaminergic neurons to a certain extent.This will provide a new experimental basis for the effect of neuroinflammation on the occurrence and development of Parkinson’s disease.