五味子甲素对K562_ADR_HL60_A_省略__MCF_7_ADR多药耐药逆

K562/ADR、HL60/ADR、MCF-7/ADR1000712010－11－192010－12－27No30973561/[e-mail][e-mail]284.1R329.24R733.73R737.9R915R979.1A1001－1978201103－0329－06schizandrinAordeoxyschizan-drinschAK562/ADR、HL60/ADR、MCF-7/ADR。MTTschAschA、-123P-gpReal-timePCRschAmdr1mRNAmrp1mRNAschAGSH。schAschA、123schAK562/ADR、HL60/ADR24hP-gpmdr1、mrp1schAK562/ADR、HL60/ADR4h。schAK562/ADR、HL60/ADRP-gpmdr1、mrp1schA。MDR1MRP1K562/ADR、HL60/ADR、MCF-7/ADRmultidrugresistanceMDR。1。2030P-gp150P-gpFDA2P-gp、3MDR。4、5、6－7、8。9。mdr1P-gpMRP1MRP1glutathioneGSHP-gp、MRP1GSHP-gpK562/ADR、MCF-7/ADRMRP1HL60/ADR。schAK562/ADR、HL60/ADR、MCF-7/ADR。11．11．1．1K562/ADR、HL-60/ADR、MCF-7/ADR。10%RPMI164037℃5%CO22～3d1。K562/ADR、MCF-7/ADR0.8mg·L－1HL-60/ADR0.5mg·L－1。2。1．1．2、、、·923·ChinesePharmacologicalBulletin2011Mar273329～34RPMI1640InvitrogenP-gpeBioscience123SigmaRealMasterMixSYBRGreen、cDNA、RNAR＆DMDR15'-CCCATCATTGCAAT-AGCAGG-3'5'-GTTCAAACTTCTGCTCCT-CA-3'157bpMRP15'-GGAC-CTGGACTTCGTTCTCA-3'MRP15'-CGTC-CAGACTTCTTCATCCG-3'292bpβ-actin5'-TGCACAACGGCTCCGGCA-3'β-actin5'-AAGGTGTGGTGCCAGATTTTC-3'。390bp。1．1．3CO2HeraeusCliniBio128OLYMPUSLDZ4-0.8FACSCaliburBectonDikinsonRe-al-timePCR。1．21．2．1MTTschAMTT。90μlK562、K562/ADR、MCF-7、MCF-7/ADR1×104HL60、HL60/ADR2×1049610μl48h5g·L－1MTT10μl37℃4h10%SDS100μl10min。550nmOD。=IC50/IC50=IC50/IC50+schA1．2．2schADNR、123P-gp5×108cells·L－12mg·L－1、2mg·L－1123Rh123schA37℃60minPBSmeanfluorescencein-tensityMFI。5×108cells·L－1schA37℃24hPBSP-gpUIC-24℃30minPBS。、P-gp488nm575nm123488nm538nm。1．2．3Real-timePCRschAmdr1、mrp1mRNA。TRIzolRNARNA。RNAcDNA37℃60min。SYBRGreenrealMasterMixReal-timePCRmdr1、mrp1β-actinPCRSYBRGreenrealMasterMix10μlddH2O7μl1μlcDNA1μl20μl。mdr1、mrp1、β-ac-tin95℃2min94℃15s56℃15s72℃30s30CT32－△△ct。1．2．4schAK562/ADR、HL60/ADRGSH1.0×109cells·L－1。4hPBS1mlPBS350HZ1500r·min－110min。450nmODODODGSH3。1．2．5珋x±stSAS9.1。22．1schAK562/ADR、HL60/ADR、MCF-7/ADRschAschA＜10%。SchAK562/ADR80μmol·l－1HL60/ADR40μmol·L－1MCF-7/ADR60μmol·L－1。2．2schAK562/ADR、·033·ChinesePharmacologicalBulletin2011Mar273HL60/ADRMTTschAP-gpK562/ADRDOX、DNR、VCR、PTXP＜0.05schAVCRPTX5-FU5-FUP-gpTab1。MRP1HL60/ADRschATab1ThereversalfoldsofschAwithDNRDOX5-FUVCRPTXfor48hinK562/ADR珋x±sn=3Concentration/μmol·L－1K562/ADRIC50/mg·L－1ReversalfoldDNRcontrol17．7±7．2－DNR+schA200．47±0．27＊＊37．7schA500．12±0．06＊＊147．5DOXcontrol28．9±12．5－DOX+schA201．92±0．94＊＊15．05schA500．38±0．26＊＊76．15-FUcontrol19．6±10．4－5-FU+schA2019．3±11．31．01schA5018．7±9．11．04VCRcontrol14．58±7．3－VCR+schA200．11±0．09＊＊132．5schA500．09±0．07＊＊162PTXcontrol1．06±1－PTX+schA200．62±0．451．7schA500．26±0．24*4*P＜0.05＊＊P＜0.01vscontrolcellsDOX、DNRP＜0.01P-gpP-gpMRP1Tab2。Tab2ThereversalfoldsofschAwithDNRDOX5-FUVCRPTXfor48hinHL60/ADR珋x±sn=3Concentration/μmol·L－1HL60/ADRIC50/mg·L－1ReversalfoldDNRcontrol13．4±5．7－DNR+schA200．91±0．43＊＊14．7schA400．30±0．17＊＊44．2DOXcontrol11．7±4．9－DOX+schA201．19±0．64＊＊9．8schA400．28±0．15＊＊41．25-FUcontrol4．9±1．6－5-FU+schA204．7±2．21．04schA404．6±1．81．06VCRcontrol13．6±2．6－VCR+schA200．57±0．23＊＊23．5schA400．21±0．10＊＊63．7PTXcontrol0．96±0．44－PTX+schA200．9±0．321．05schA400．86±0．271．1＊＊P＜0.01vscontrolcellsFig1Dose-dependenteffectsoftheschAontheintracellularDNRaccumulationinMDRcell珋x±sn=3*P＜0.05＊＊P＜0.01vscontrolcells·133·ChinesePharmacologicalBulletin2011Mar2732．3schADNRDNR2mg·L－1DNRschAK562/ADR、HL60/ADR、MCF-7/ADR37℃60minschADNRschADNRP-gpMRP1Fig1。2．4schA123Rh123K562/ADR、MCF-7/ADRRh123Rh123P-gp5μmol·L－1Rh123schA60minschAP-gpK562/ADR、MCF-7/ADRRh123schAP-gpschA60minschAP-gpP-gpFig2。Fig2Dose-dependenteffectsoftheschAontheintracellularRh123accumulationinK562/ADRMCF-7/ADRcellsbyflowcytometry珋x±sn=3＊＊P＜0.01vscontrolcells2．5SchAP-gpK562/ADRP-gpschAK562/ADRschAP-gpFig3。Fig3Dose-dependenteffectsoftheschAontheexpressionofP-gpinK562/ADRcellsbyflowcytometry珋x±sn=3*P＜0.05＊＊P＜0.01vscontrolcells2．6SchAK562/ADRmdr1HL60/ADRmrp1mRNAschAK562/ADR、HL60/ADR24hReal-timePCRschAmdr1、mrp1Fig4。2．7SchAK562/ADR、HL60/ADRGSHschAK562/ADR、HL60/ADRGSHschAHL60/ADRMRPGSH－。Tab3Dose-dependenteffectsoftheschAontheGSHcontentinK562/ADRHL60/ADRcellsbyhumanGSHELISAkit珋x±sn=3GroupConcentration/μmol·L－1GSHConcentration/U·L－1K562/ADRHL60/ADRControl－187．2±16．3193．0±14．2SchA10－167．3±21．620174．6±23．4166．4±25．340－144．6±30．5＊＊50156．3±14．1－80137．1±26．5＊＊－＊＊P＜0.01vscontrolcells3MDR。。·233·ChinesePharmacologicalBulletin2011Mar273Fig4Dose-dependenteffectsoftheschAontheexpressionofMDR1MRP1inK562/ADRHL60/ADRcellsbyReal-timePCR珋x±sn=3*P＜0.05＊＊P＜0.01vscontrolcellsschAMDRMTTschAK562/ADR、HL60/ADRVCR、PTX、DNR、DOX、5-FUschAP-gp、MRP1。schAP-gp、MRP1P-gp、MRP1。schAK562/ADR、HL60/ADRGSHschAGSHGSHGSHMRP1。schA。1OzaAM．ClinicaldevelopmentofP-glycoproteinmodulatorsinon-cologyJ．NovartisFoundSymp2002243103－15．2PeerDMargalitR．FluoxetineandreversalofmultidrugresistanceJ．CancerLett20062372180－7．3LiXJZhaoBLLiuCTetal．Scavengingeffectsonactiveoxy-genradicalsbyschizandrinswithdifferentstructuresandconfigura-tionsJ．FreeRadicBiolMed19909299－104．4．HIV-1J．20064140－4．4TianRRXiaoWLYangLMetal．Theisolationofrubrifloralig-nanAanditsanti-HIV-1activitiesJ．ChinJNatMed20064140－4．5KuoYHKuoLMChenCF．Fo