Shvoong首页 > 医药 & 健康 > Effect of Astragalus Membranaceus on Ca ̄(2+)Influx and CVB3 RNA Replication in Cultured Neonatal Rat

.

Effect of Astragalus Membranaceus on Ca ̄(2+)Influx and CVB3 RNA Replication in Cultured Neonatal Rat

摘要撰写人 : TsingHua
浏览次数 : 13  词语: 300   出版日期: 三月 20, 1996
EffectofAstragalusMembranaceusonCa ̄(2+)Influ xandCVB3RNAReplicationinCulturedNeonatalRat HeartCellsInfectedwithCVB3GUOQi(郭棋);PENGTia n-oing(彭天庆);YANGYing-zhen(杨英珍)(ShanghaiInst ituteofCardiovascularDiseases,ZhongShanHosp italofShanghaiMedicalUniversity,Shanghai200 032)GUQuan-bao(顾全保)andZHAOJian-xing(赵剑星)(Sh anghaiInstituteofCellBiology,AcademiaSinica ,Shanghai200031)ABSTRACTHerewasinvestigated theeffectofRadixAstragalusMembranaceusIAM)o nCaz+influxacrossthemyocardialplasmamembran eandcoxsackievirusB3(CVB3)-RNAreplicationin culturedneonatalratheartcellsinfectedwithCV B3.ItwasfoundthattheOa2+intluxcouldbeinhibi tedsigniticantlybyAMbothinheartcellsintecte dwithCVB3for48hoursandinnormalcontrolheartc ells.Inaddition.theCaz+intluxandtheamountso fCVB3-RNAinrnyocytessimultaneouslyintectedw ithCVB3andtreatedwithAMfor48hourswerestatis ticallydecreasedcomparedwiththatinCVB3-infe ctedcontrOIcells.Thesephenomenasug-gestedth atAMcouldexerttheeffectsofdecreasingtheseco ndaryCa2+damages,irnprovingtheabnormalmyoca rdialelectricactivityandinhibitingreplicati onofCVB3-RNAinmyocardium.Thus,itisarational choicetotreatpatientswithAMinviralmyocardit is.KEYWORDSCoxsackievirusB3,viralmyocarditi s,Ca2+influx,RadixAstragalusmembranaceusCVB infectionisbelievedtobeassociatedwithmosthu manviralmyocarditis.Itisgen-erallyknownthat viralreplicationinmyocardi-umresultingininf lammatorymyocardialnecrosisandvirusmediated cellularimmunityinducingmyocardialinjuryare mechanismofpathogenesisofviralmyocarditis,b othdiseaseprocessesapttocauseCa2+overloadof heartcells,aggravatingthemyocardialdam-ages (1-4).ItwasfoundthatRadixAstragalusmem-bran ceus(AM)couldpreventmyocardialdam-agefromCV Binfectionandimprovetheelec-tricalactivitie sofCVB-infectionmv-ocytes(5,6).Inthisstudy, theeffectofAMonCa2+influxacrossthemyocardia lmembraneandcoxsackievirusB3(CVB3)-RNArepli cationinculturedneonatalratheartcellsinfect edwithCVB3wasinvestigatedtoascertainitsthev alueforclinicalapplication.METHODSPreparati onotCulturedHeartCellsandCVB3InoculationSin glecellsuspensionsofventriclesfromnewbornSp rague-Dawleyratswerepreparedbytheprocedures describedpreviously(4),andmaintainedingrowt hmediaincluding20%newbornbovineserum,1%L-gl utamine(200mmol/L),penicillin(100g/ml)andst repto-mycin(100g/ml)inEagle'sminimumessen-t ialmedium(MEM).Afterincubationat37for24hour s,thecells(5105cellsin1mlgrowthmediaforeach well)weredividedintotwogroups:infectedgroup anduninfectedgroup.Afterremovingthesupernat ant,0.5mlofgrowthmediacontainning100TCID-50 (tissueinfectiousdose-50)ofCVB3(Nancystrain )wasaddedtothecellsoftheinfectedgroup,andon lygrowthmediawasaddedtothoseofthetouninfect edgroup.Onehouraftervirusabsorption,super-n atantsinwellswereremovedand1mlofgrowthmedia wasaddedtoeachwell,incu-batedat37againforfo llowingexperiments.AMTreatmentandCa2+Intlux DeterminationForty-eighthoursafterviruschal lenge,thecellswerewashedthreetimeswithCa2+- freeRinger'ssolutionanddividedintofourgroup s:uninfectedcontrol,infectedcontrol,AMcontr olandinfected-Amtreatedgroup.Twohundredtilo fRinger'ssolutioncontaining50mol/L[Ca2+]ope rwellwasaddedtoeverygroup,whileadditional0. 1mlofAM(4g/2mlherb/ampule,840308,suppliedby PharrnaceuticalDepartmentofShanghaiZhongsha nHospital)perwellwasaddedtothelattertwogrou ps.Aftercellswereincubatedat37for5min.,thes upernatantswereremovedand250PIofRinger'ssol ution-labelledwith37kBq/mlof45Ca2+(1Ci,supp liedbyDuPontCom-pany,USA)wasaddedtoeachwell (alsocon-tainning0.2g/mlAMinthelattertwogro ups).Theplateswereincubatedat37and45Ca2+-la belledsolutionwasdischargedat1,5and10minres pectively.After3fastwashesofthecellstimeswi th7%sorbitolso-lutioncontainning1mmol/LLaon ice,0.5mlof5%sodiumdodecylsulphate(SDS)wasa ddedtoeachwellandkeptfor2hoursatrcomtempera ture.Twohundredlofhomog-enizedsuspensionfro meachwellwascountedforCPMonaliquidscintillo meter(BECK-MANLS6500).Theaveragecontentofpr oteinineachwellwasdeterminedbylowry'smethod .TheCa2+-influxacrossthemyocardialplasmamembranewasexpressedasfollows:{cpm[Ca2+]o(nmol)}/{totalcpmprotein(mg)}(4).Thesameexperimentaldesignforcellswith48hoursafterviruschal

More abstracts about the Effect of Astragalus Membranaceus on Ca ̄(2+)Influx and CVB3 RNA Replication in Cultured Neonatal Rat
请给本文打分 : 1 2 3 4 5


标记

评论

Read Free Summaries - Write and Get Paid

Summarize Human Knowledge on Shvoong. Join us!

------