Study On Neuroprotective Effect Of Blocking Pannexin1 Channel On Parkinson’s Model Mouse

Parkinson’s Disease(PD)is a common neurodegenerative disease.According to statistics from the World Parkinson’s Association,there are close to 6 million PD patients in the world,and PD patients in China account for about half.It is still increasing at a rate of one hundred thousand every year,which puts huge pressure on our country’s social public health and medical resources.The pathological features of PD are the appearance of characteristic eosinophilic inclusions Lewy bodies and the degeneration and loss of dopaminergic neurons in the Substantia Nigra(SN).Symptoms in the PD patient include motor symptoms such as tremor,muscle stiffness,and other movement disorders,as well as non-motor disorders such as constipation,depression,which seriously affect the normal life of the patient.At present,the specific pathogenesis of PD is still unclear.The potential pathogenesis includes apoptosis,neuroinflammation,mitochondrial dysfunction,oxidative stress,etc.The most effective treatment for patients with early PD is to use levodopa(L-DOPA),dopamine receptor agonists and other drugs for treatment,but long-term use of these drugs will lead to weakened efficacy and severe side effects.In addition,some surgical treatments for patients with advanced PD(such as deep brain stimulation)can cause brain microhemorrhage and other injuries.Therefore,finding new drugs or targets for PD treatment is of great significance to the treatment of PD.Pannexin1(Panx1)channel is a newly discovered iron channel that mediates a variety of important physiological functions,such as: blood flow regulation,neuroinflammatory response.Furthermore,the Panx1 channel is involved in a variety of neurological diseases,such as cerebral ischemia,epilepsy.But so far,the role of Panx1 channel in PD is still unclear.Through experiments,we found that the expression of Panx1 in the SN of PD model mice was significantly higher than that of control mice.Combined with the Panx1 channel can mediate neuroinflammatory response,we believe that Panx1 channel may be related to PD.Through behavioral and biochemical experiments,it was found that blocking the Panx1 channel with CBX can significantly improve the dyskinesia and typical pathological symptoms of PD model mice.This subject have studied the neuroprotective effect of CBX blocking Panx1 channel on PD model mice and its possible mechanism,providing new ideas and targets for the treatment of PD.The research content of this paper is as follows:1.Using MPTP to induce the establishment of chronic and subacute PD models.Those two models and each control group were compared through two behavioral experiments of open field and rotarod test,and TH immunohistochemistry.The results showed that the motor symptoms and pathological characteristics of subacute PD mice were more obvious.Therefore,the subacute PD model mice were finally selected as the research object;2.C57BL/6 mice were randomly divided into three groups: control group(Control),subacute PD model group(MPTP),and CBX treatment group(CBX+MPTP).Control group: injected the same amount of saline for 7 consecutive days;model group: injected MPTP for 7 consecutive days;CBX treatment group:received panx1 classic blocker CBX(Carbenoxolone)half an hour before intraperitoneal injection of MPTP for 7 consecutive days;Through four kinds of animal behavior experiments,exploring the effect of blocking panx1 channel on the exercise ability of PD model mice;Detecting the expression of TH in the midbrain SN of mice by Western blot,and using immunohistochemistry to observe the changes in pathological characteristics of PD mice,explore the effect of blocking the panx1 channel on the pathological characteristics of PD model mice.The results showed that compared with the model group of mice,the dyskinesia of the CBX treatment group was significantly improved and the number of TH-positive cells and neurons in the midbrain SN was significantly increased,indicating that CBX blocking the Panx1 channel has neuronal protection in PD mice effect.3.To explore the possible mechanism of blocking the Panx1 channel on the treatment of PD model mice,the expression levels of NLRP3,ASC,and caspase1 in the SN of mice from the three groups were detected by western blot.Subsequently,the content of TNF-α and IL-1β in the SN of mice from the three groups were detected by ELISA.The results showed that the expression of NLRP3,ASC,and caspase1 protein in PD model mice were significantly increased,comparing with control group.After CBX blocked the panx1 channel,the expression of NLRP3,ASC,and caspase1 were all down-regulation;ELISA test showed that CBX can significantly reduce the level of pro-inflammatory cytokines in PD model mice.We believe that the panx1 channel mediate the activation of the inflammasome,triggers a neuroinflammatory response,and affects PD model mice.Blocking the panx1 channel has a protective effect on PD mice.In summary,blocking the Panx1 channel by CBX can inhibit the NLRP3-ASC-caspase1 signaling pathway,and reduce the level of neuroinflammation,having neuroprotective effects on PD model mice.These studies provide new ideas and new targets for PD treatment.