内毒素休克小鼠肝脏线粒体差异蛋白质组的鉴定

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1、硕士学位论文before2-Delectrophoresis，andthenseparateuptothreedifferentproteinsamplesatthesametimeinonetwo．dimensionalgel．Theapplicationoftheinternalstandardcouldfurtherincreasethecredibilityoftheexperiment，andensuretheresultscouldreflecttherealbiologicaldifferences，

2、whileavoidinfluenceofsystematicerrorsonexperimentalresults．SinceThemostobviousadvantageofDIGEsystemisintegratingtheadvantagesofbothCyDyeDIGEdyemultiplelabelingmethodandDeCyderdifference2-Danalysissoftware．DeCydersoftwaretakest11eadvantageofthespotsCO·detecting

3、algorithm，whichCanautomaticallydetectedfluorescenceimages，eliminatebackground，quantifiy,normalizeandmatchspotsingel，thus，systematicerrorscausedbydifferentoperatorsconbeeliminated．BasedontheadvantagesofDIGEtechnologyplatform，greatopportunitiescomestoustodiscove

4、rtheinformationofdifferentialproteomecontainedinmicelivermitochondria、析tllendotoxicshockandprovidenoveltheoriesforsepsisresearch．Atechnologyplatformbasedondifferencegelelectrophoresis，MALDITOF／TOFmassspectrometryandbioinformaticsanalysiswasestablished缶st，andth

5、ensomeassaysonitssensitivityandreproducibilitywasdoneinourresearch．AfterduplicatingtheendotoxicshockmodelofBALB／cmiceafteranesthesia,wecollectedmiceliver,applieddifferentialcentrifugationanddensitygradientcentrifugationmethodtoextractpurifiedlivermitochondfia,

6、andextractedtheentiremitochondrialproteinsbylysisbuffer．ThenwegottheexpressionpatternsoflivermitochondrialdifferentialproteomeofendotoxicshockmicebyusingDIGEtechnology．ThroughanalysisbyuseofDeCyderdifference2一Danalysissoftware，26differentialproteinspotswerefou

7、nd．，nlenumbersofup—regulatedanddown-regulatedproteinsareboth13．Finally，atotalof22proteinshavebeenidentifiedamongthesedifferentialproteinspotsbyusingmatrix-assistedlaserdesorptionionizationmassspectrometry(MALDI—TOF／TOF)．neCO．expressionofthesedifferentialprotei

8、nsindicatesthattheymusthavedirectorindirectrelations．Therefore，howtofindoutanddiscoverthepathophysiologymeaningoftheserelationsinendotoxicshocklivermitochondriaisthekeyforfurtherre