p75神经营养蛋白受体基因敲除对小鼠面神经损伤后神经再生的影

28120102WestChinaJournalofStomatologyVol．28No.1Feb.2010http//[]2009-01-13[]2009-11-21[]Y2008C54[]1965—[]Tel[phone][]1000－1182201001－0095-042tyrosineproteinkinase,TrkTrkA、TrkB、TrkCp75p75neurotrophinreceptorp75-NTR。p75NTR[1]p75NTR[2]。p75NTR[3]p75NTRp751211（1.；2.，250012）[]p75p75NTR。p75NTR129sv2mm2BCTB3、7d。4FastBlue。p75NTR129svP＜0.05。p75NTRP＜0.01。p75NTR。[]p75B[]Q786[]A[doi]10.3969/j.issn.1000-1182.2010.01.025Influenceofp75neurotrophinreceptorknockoutontheregenerationoffacialnervesaftercrushinjuryinmouseZHANGFeng-he1,HUANGPing2,YANGPi-shan1ZHANGXue1.1.Dept.ofOralandMaxillofacialSurgery,SchoolofStomatology,ShandongUniversity,Jinan250012,China;2.Dept.ofRadiotherapyQiluHospital,ShandongUniversity,Jinan250012,China[Abstract]ObjectiveToinvestigatetheroleofp75neurotrophinreceptorp75NTRintheregenerationoffacialnervecrushinjury.MethodsInp75NTRknockoutmiceandwildtypemice,theregeneratingfibresinthefacialnervewerealsolabelledbyananterogradetracercholeratoxinBCTB.Thenextdayafterinjuryoffacialnerve,CTBwasinjectedintothetrunkofthenerveintheproximalsideofthecrush,andthenanterogradetracingandimmunohistochemistrywereusedtoexaminetheregenerationofaxonsafterfacialnervecrushinjury.Inp75NTRknockoutmiceandwildtypemice,thefacialnervesononesidewerecrushedandregeneratingneuronsinthefacialnervenucleuswerelabelledbyFastBlue.Thefacialnervetrunkwascutinthebifurcatedregioninthe4thdayafterinjuryandthestumpwasinsertedintoasmallpolymertubecontainingFastBlue.Retrogradetracingandlablingmotoneuroncountingwereusedtoexaminethesurvivalofmotoneuronsinthefacialnervenucleusafterfacialnervecrushinjury.ResultsTheresultsshowedthattheaxonalgrowthofinjuredaxonsinthefacialnerveofp75NTRknockoutmicewassignificantlyretarded.Thenumberofregeneratedneuronsinthefacialnervenucleusinp75NTRknockoutmicewassignificantlyreducedP＜0.05.Immunohistochemicalstainingofregeneratingaxonsalsoshowedthereductioninnerveregenerationinp75NTRknockoutmiceP＜0.01.Conclusionp75NTRplaysanim-portantroleintheregenerationofinjuredperipheralnervesafterinjury.[Keywords]p75neurotrophinreceptorcholeratoxinBregenerationknockout95··95··28120102WestChinaJournalofStomatologyVol．28No.1Feb.2010http//。p75NTRp75NTRp75NTR。11.190129svp75NTR45。6～825～29g。2BcholeratoxinBCTB355FastBlue10。1.2OlympusSZX-12OlympusLeicaLeicaHamiltonCTB、FastBlue、43GAP43SigmaCTBListBiologicals-PGP9.51∶5000Invitrogenrabbitanti-calcitoningenerelatedpeptideCGRPMerke。1.31.3.11μL·g-10.5μL·g-1illumxylazil901cm2mm30s90°30s。。0/8。1.3.24129svp75NTR105%FastBlue0.2μL。。74%4%24h10%、20%、30%-20℃16μmPBST。1.3.3370nm。311。1.3.42129svp75NTR5CTB0.2μL1min。34%。4%24h10%、20%、30%-20℃20μmCTB。1.3.53d7dPBST20%normalhorseserumNHS2h。CTB1∶10000、GAP-431∶500、-PGP9.51∶5000CGRP。PBSTFITC1∶1002h。CTB。GAP43、PGP9.5CGRP3mm3。。1.4SPSS12.0t。22.1p75NTRCTB。3d129sv2mm。3d6mm。p75NTR1mm3d2.5mm。2.2p75NTR7dp75NTR129sv96··96··28120102WestChinaJournalofStomatologyVol．28No.1Feb.2010http//。129sv77.8±4.5%p75NTR53.1±5.2%P0.05。A、B129sv、C、Dp75NTR、。1129svp75NTR×100Fig1Theregenerationoffacialmotoneuronsinthefacialnervenucleusinwildtype129svmiceandp75NTRknockoutmicefluorescencemicroscope×100A、C、Ep75NTRGAP43、PGP9.5、CGRPB、D、F129svGAP43、PGP9.5、CGRP。2129svp75NTRGAP43、PGP9.5、CGRP×400Fig2ThefacialnervefibresimmunoreactiveforGAP43,PGP9.5andCGRPinwildtype129svmiceandp75NTRknockoutmicefluoresce-ncemicroscope×4002.3p75NTRGAP43、PGP9.5CGRP23mm2。129svGAP43、PGP9.5CGRP27.5±3.2、38.6±4.1、20.4±2.9。p75NTRGAP43、PGP9.5CGRP14.2±2.2、20.8±3.4、9.7±2.1P0.01。3p75NTR。Boyd[4]p75NTR。p75NTR[5]。p75NTR。p75NTR。97··97··28120102WestChinaJournalofStomatologyVol．28No.1Feb.2010http//。p75NTR。CGRP、PGP9.5GAP43p75NTRp75NTR。p75NTR。p75NTR[6]。p75NTR。p75NTR[7]。p75NTRp75NTR。p75NTR。p75NTRthereceptor-mediatedintramembraneproteolysisRIP[8-9]。RIP[10]。p75NTRRIP。p75NTR。p75NTR。p75NTRNgR/LINGO-1/p75NTR[11]。p75NTRNgR/LINGO-1/p75NTR。p75NTRp75NTR。p75NTR。。[12]。p75NTR[13]。p75NTRp75。。[][1]SongXY,ZhongJH,WangX,etal.Suppressionofp75NTRdoesnotpromoteregenerationofinjuredspinalcordinmice[J].JNeurosci,2004,242542-546.[2]SongXY,ZhouFH,ZhongJH,etal.Knockoutofp75NTRimpairsre-myelinationofinjuredsciaticnerveinmice[J].JNe-urochem,2006,963833-842.[3]JohnsonEMJr,TaniuchiM,DiStefanoPS.Expressionandpos-siblefunctionofnervegrowthfactorreceptorsonSchwanncells[J].TrendsNeurosci,1988,117299-304.[4]BoydJG,GordonT.Adose-dependentfacilitationandinhibitionofperipheralnerveregenerationbybrain-derivedneurotrophicfactor[J].EurJNeurosci,2002,154613-626.[5]ScottAL,BorisoffJF,RamerMS.DeafferentationandneurotrophinmediatedintraspinalsproutingAcentralroleforthep75neu-rotrophinreceptor[J].EurJNeurosci,2005,21181-92.[6]FunakoshiH,FrisénJ,BarbanyG,etal.DifferentialexpressionofmRNAsforneurotrophinsandtheirreceptorsafteraxotomyofthesciaticnerve[J].JCellBiol,1993,1232455-465.[7]ZhouXF,LiHY.Rolesofglialp75NTRinaxonalregeneration[J].JNeurosciRes,2007,8581601-1605.[8]AhmedZ,MazibradaG,SeabrightRJ,etal.TACE-inducedcleavageofNgRandp75NTRindorsalrootganglionculturesdisinhibitsoutgrowthandpromotesbranchingofneuritesinthe