Primary Research Of TSPO After Chronic Sleep Deprivation Of Mice And Effects Of Its Ligand Etifoxine On NLRP3 Inflammasome

Objectives:Observe the effects of chronic sleep deprivation on protein expression and gene expression of TSPO,coexpression of TSPO with glias cells in the hippocampus of mice and the effects on protein expression of NLRP3 inflammasome.Further explore the effects of TSPO ligand etifoxine on protein expression of NLRP3 inflammasome and gene expression level of inflammatory factors in chronic sleep deprivation.To explore the relationship between TSPO expression and inflammatory response and the neuroprotective effect of its ligand after chronic sleep deprivation,which provides a possible idea for in vivo imaging of chronic sleep deprivation and in vivo monitoring the response of neuroinflammatory,and provides a new method for comprehensive treatment of chronic sleep deprivation.Method:Male C57BL/6J mouses were randomly divided into four groups which 12 mice in each group:CC group?SD group?SD+Etifoxine?SD+Vehicle.All mice fed in suitable environmen,SD group:7 days of chronic sleep deprivation with MMPM;SD+Etifoxine:After chronic sleep deprivation,given etifoxine(dissolved in 1% Tween 80 in 0.9%NaCl solution)at the dose of 50mg/kg by i.p.injection at indicated time for three days;SD+Vehicle:After chronic sleep deprivation,received an equal volume of vehicle(0.9%NaCl solution containing 1%Tween 80)for three days.Immunofluorescence staining was used to examine TSPO~+cells in mouse hippocampus,Double-labled immunofluorescence assese the coexpression of TSPO and glias.Western blots analyzed for TSPO and NLRP3 inflammasome components,the gene expression of NLRP3?IL-6?IL-1??IL-18?TNF-?was extracted from brain tissue in mouse hippocampus for real-time polymerase chain reaction analysis.Results:1?We also observed the amount of TSPO positive cells?protein and mRNA were significantly larger in mouse hippocampus of chronic sleep deprivation than in blank control.(P<0.01?P<0.001?P<0.01 respectively).2?Double-labeled immunofluorescence shows TSPO~+/Iba1~+and TSPO~+/GFAP~+cells in brain slices of mouse.The expression of TSPO was mostly seen in Iba1~+cells.Compared with CC group,TSPO~+/Iba1~+and TSPO~+/GFAP~+cells increased in SD group(P<0.01?P<0.01 respectively).3?Compared with CC group,the expression of NLRP3?Caspase-1?IL-1?protein increased in SD group(P<0.01?P<0.01 P<0.01 respectively),the gene expression of NLRP3 and inflammatory factors(IL-6?IL-1??IL-18?TNF-?)increased significant in SD group(P=0.015?P<0.01?P=0.028?P=0.017?P<0.01 respectively).4?Compared with SD group,the expression of NLRP3?Caspase-1?IL-1?protein and the gene expression of inflammatory factors had no significant difference in SD+Vehcile group.5?Compared with SD group,the expression of NLRP3?Caspase-1?IL-1?protein decrease in SD+etifoxine group(P=0.031,P=0.029,P<0.01 respectively),the gene expression of NLRP3 and inflammatory factors(IL-6?IL-1??IL-18?TNF-?)decrease significant in SD+etifoxine group(P=0.023?P=0.035?P=0.023?P=0.044?P=0.049).Conclusions:1?The expression of TSPO was mostly seen in astrocytes and microglias.The expression of TSPO gene and protein increased in hippocampus of mice after chronic sleep deprivation.It provides a possible idea for in vivo imaging of chronic sleep deprivation.2?TSPO may be involved in the activation of NLRP3 inflammasome after chronic sleep deprivation,which may be one of the mechanisms for neuroinflammation caused by chronic sleep deprivation.3?TSPO ligand inhibited neuroinflammation,which may play a protective role in neuroinflammation caused by chronic sleep deprivation.Targeted TSPO therapy may be an option for the comprehensive treatment of patients with chronic insomnia.4?Etifoxine,as a non-benzodiazepine anti-anxiety drug,may improve inflammatory injury in patients with chronic insomnia,which provides the clinical comprehensive treatment of chronic insomnia with anxiety and depression.