简介：AIM:ToinvestigatethepreciserolesofCARinCCl4-inducedacutehepatotoxicity.METHODS:Toprepareanacuteliverinjurymodel,CCl4wasintraperitoneallyinjectedinCAR+/+andCAR-/-mice.RESULTS:ElevationofserumalanineaminotransferaseandextensionofcentrilobularnecrosiswereslightlyinhibitedinCAR-/-micecomparedtoCAR+/+micewithoutPB.AdministrationofaCARinducer,PB,revealedthatCCl4-inducedlivertoxicitywaspartiallyinhibitedinCAR-/-micecomparedwithCAR+/+mice.Ontheotherhand,androstanol,aninverseagonistligand,inhibitedhepatotoxicityinCAR+/+butnotinCAR-/-mice.Thus,CARactivationcausedCCl4hepatotoxicitywhileCARinhibitionresultedinpartialprotectionagainstCCl4-inducedhepatotoxicity.TherewerenodifferencesintheexpressionofCYP2E1,themainmetabolizingenzymeforCCl4,betweenCAR+/+andCAR-/-mice.However,theexpressionofotherCCl4-metabolizingenzymes,suchasCYP2B10and3A11,wasinducedbyPBinCAR+/+butnotinCAR-/-mice.AlthoughthemainpathwayofCCl4-inducedacuteliverinjuryismediatedbyCYP2E1,CARmodulatesitspathwayviainductionofCYP2B10and3A11inthepresenceofactivatororinhibitor.CONCLUSION:ThenuclearreceptorCARmodulatesCCl4-inducedliverinjuryviainductionofCCl4-metabolizingenzymesinthepresenceofanactivator.OurresultssuggestthatdrugsinteractingwithnuclearreceptorssuchasPBmightplaycriticalrolesindrug-inducedliverinjuryordrugdruginteractioneventhoughsuchdrugsthemselvesarenothepatotoxic.

简介：AIM:Toinvestigatetheanti-tumoreffectsofnuclearfactor-κB(NF-κB)inhibitorSN50andrelatedmechanismsofSGC7901humangastriccarcinomacells.METHODS:MTTassaywasusedtodeterminethecytotoxiceffectsofSN50ingastriccancercelllineSGC7901.Hoechst33258stainingwasusedtodetectapoptosismorphologicalchangesafterSN50treatment.Activationofautophagywasmonitoredwithmonodansylcadaverine(MDC)stainingafterSN50treatment.Immunofluorescencestainingwasusedtodetecttheexpressionoflightchain3(LC3).MitochondrialmembranepotentialwasmeasuredusingthefluorescentprobeJC-1.Westernblottinganalysiswereusedtodeterminetheexpressionofproteinsinvolvedinapoptosisandautophagyincludingp53,p53upregulatedmodulatorofapoptosis(PUMA),damage-regulatedautophagymodulator(DRAM),LC3andBeclin1.Wedetectedtheeffectsofp53-mediatedautophagyactivationontheapoptosisofSGC7901cellswiththep53inhibitorpifithrin-α.RESULTS:TheviabilityofSGC7901cellswasinhibitedafterSN50treatment.Inductionsintheexpressionofapoptoticproteinp53andPUMAaswellasautophagicproteinDRAM,LC3andBeclin1weredetectedwithWesternblottinganalysis.SN50-treatedcellsexhibitedpunctuatemicrotubule-associatedprotein1LC3inimmunoreactivityandMDC-labeledvesiclesincreasedaftertreatmentofSN50byMDCstaining.CollapseofmitochondrialmembranepotentialΔψweredetectedfor6to24hafterSN50treatment.SN50-inducedincreasesinPUMA,DRAM,LC3andBeclin1andcelldeathwereblockedbythep53specificinhibitorpifithrin-α.CONCLUSION:Theanti-tumoractivityofNF-κBinhibitorsisassociatedwithp53-mediatedactivationofautophagy.

简介：AIMToexploretheprotectiveeffectsandunderlyingmechanismsoftotalpolysaccharidesoftheSijunzidecoction(TPSJ)ontheepithelialbarriersinvitro.METHODSCaco-2cellmonolayersweretreatedwithorwithoutTPSJinthepresenceorabsenceofTNF-α,andparacellularpermeabilityandtransepithelialelectricalresistance(TEER)weremeasuredtoevaluatetheepithelialbarrierfunction.Immunofluorescenceandwesternblottingwererespectivelyusedtoevaluatethedistributionandexpressionofthetightjunctionproteinsclaudin1,claudin2,zo3,andoccludininCaco-2cells.Westernblottingwasalsousedtoevaluatethecellularexpressionofmyosinlightchain(MLC),phosphorylatedMLC(pMLC),MLCkinase(MLCK),andnuclearfactor(NF)-κBp65.RESULTSTPSJpromotedtheproliferationofCaco-2cellsandinhibitedTNF-α-inducedsecretionofpro-inflammatorycytokines.Furthermore,TPSJsignificantlyamelioratedboththereductionofTEERandtheincreasedparacellularpermeabilityobservedintumornecrosisfactor(TNF)-α-damagedCaco-2monolayers.Furthermore,TPSJremarkablyattenuatedTNF-α-inducedmorphologicalchanges,downregulatedtheexpressionofclaudin1,claudin2,zo3,andoccludin,andmarkedlysuppressedTNF-α-mediatedupregulationofp-MLCandMLCKexpression.Finally,TPSJinhibitedtheactivationandexpressionofNF-κBp65.CONCLUSIONOurresultsdemonstratethatTPSJalleviatestheTNF-α-inducedimpairmentoftheintestinalepithelialcellbarrierfunctionbysuppressingNF-κBp65-mediatedphosphorylationofMLCKandMLC.