克百威与DNA之间作用的光谱学研究

25,5Vol.25,No.5,pp739-74220055SpectroscopyandSpectralAnalysisMay==============================================================================,2005DNA1,1,2,1,1*1.,1000942.,100094ctDNA。,ctDNA;ctDNA。ctDNA。。;;ctDNA;;;:R931.6:A:1000-0593(2005)05-0739-04:2003-11-18,:2004-03-06:,(8000001):,1974,*(DNA)。DNA。,、DNA,[1],,DNA,[2]。DNA,DNA。DNA[3]。(Carbofuran),,25℃700µg·mL-1;、,,,、[4]1。Fig.1Structureofcarbofuran,。[5]。,;-,mRNA[6],ctDNA。11.1F-4500;Tu-1901;PHS-3。DNA(CalfThymusDNA,ctDNA),250µg·mL-1,A260/A280=1.99,260nm(ε=6600L·mol-1·cm-1),4℃;50µg·mL-1(λex=233nm,λem=316nm);EDTA-Tris,pH7.00;,。1.210mL,ctDNA,,,、;10mL、ctDNA,,,。10mL、,ctDNA,,,,,。22.1ctDNADNA,,DNADNA[7]。Fig.2TheeffectofontheUVspectrumofcarbofuranc(ctDNA)/(φ),1→6(0,10,20,30,40,50)2ctDNA,ctDNA,ctDNA[8]。2.2ctDNA3ctDNA,。ctDNA;ctDNA[9]。Fig.3ThefluorescentquenchingofctDNAtocarbofuranat37℃c(ctDNA)/(φ),1→8(0,10,20,30,40,50,60,70)2.3ctDNAn,nP+QPnQQ,P,PnQ,KK=[PnQ]/[P]n[Q][P0][P][P0]=n[PnQ]+[P],,[P]/[P0]=F/F0lg[(F0-F)/F]=lgK+nlg[Q](1)(1),(4)。Fig.4Thequenchingfunctionoflg[(F0-F)/F]-lg[Q]atdifferenttemperature4,K。F0/F=1+Kqτ0[Q]=1+Ksv[Q]。Ksv≈4～5×103L·mol-1,τ0≈10-8s[10],Kq≈4～5×1011L·(mol·s)-1。Kq≈2.0×1010L·(mol·s)-1[10],Kq,[11];,,ctDNAn,。,,ctDNActDNA-[12];,ctDNAB[12],,。2.4ctDNA-5,。,[13];,,[14]。Fig.5Theeffectofethanolonthefluorescentspectrumofcarbofuranc(ethanol)/(φ),1→6(0,10,20,30,40,50)047256,ctDNA-。,;,ctDNA,ctDNA[15],。。Fig.6TheeffectofethanolonthefluorescentspectrumofcarbofuranandctDNAc(ethanol)/(φ),1→6(0,10,20,30,40,50)2.5,DNADNA[16],:,DNA;,[17]。(1)。1ctDNA,ctDNA,ctDNA[18]。Table1Theeffectofsodiumchlorideonthefluorescentquenching[NaCl]%0.00y=1.0738x+4.01670.90y=1.0597x+3.96113(1)ctDNActDNA;ctDNA。(2)ctDNActDNA-。(3)ctDNA,ctDNA。1475[1]YANGMing().JournalofBeijingMedicalUniversity(),1998,30(2):97.[2]HUANGJin-xiang().OccupationalHealthandEmergencyRescue(),1996,14(2):52.[3]QIShu-min().ProgressofPharmacology(),1985,12:319.[4]CHENWan-yi,ZHAOZhong-hua,XUEZhen-xiangetal(,,).ProductionandSynthesisofPesticides().Beijing:ChemicalIndustryPress(:),2000.134.[5]SONGHong-yu,WANGJie(,).PesticidesScienceandMinistratioo(),2001,22(2):23.[6]LDong-yang,LIWen-lan(,).JournalofHarbinUniversityofCommerceNaturalSciencesEdition(·),2003,19(1):22.[7]YEYong,HUJi-ming,YINXin-qiang(,,).JournalofInstrumentalAnalysis(),2000,19(1):38.[8]ZHANGPeng,PANGMei-li,MENGJi-benetal(,,).ChemicalJournalofChineseUniversities(),2002,23(8):1531.[9]KumarCV,EmmaHAsuncion.J.Am.Chem.Soc.,1993,115:8549.[10]ZHANGHai-rong,GUOSi-yuan,LILinetal(,,).SpectroscopyandSpectralAnalysis(),2001,21(6):830.[11]TANGShi-hua,HUANGJian-bin(,).ActaPhys.Chem.Sinica(),2001,17(10):875.[12]WANGLei,YANGGuang,WUJian-zhongetal(,,).ActaScientiarumNaturalIumuniversitatisSunyatseni(·),1997,36(5):53.[13]LIUYi-qing,PANZu-ting,ZHANGTaoetal(,,).ChineseJournalofAnalysisLaboratory(),2002,21(1):61.[14]McraeEG.J.Phys.Chem.,1957,61:569.[15]LILai-sheng,HUANGWei-dong,YANYuan(,,).SpectroscopyandSpectralAnalysis(),2001,21(5):685.[16]RouzinaL,BloomfiedVA.J.Phys.Chem.,1996,100:4301.[17]KumarCV,TurrierRS,AsuncionEH.J.Photochem.Photobiol.A.Chem.,1993,74:235.[18]CAOYing,LIYi-jun,HEXi-wenetal(,,).ActaPharmaceuticaSinica(),1999,34(4):278.TheMechanismofCarbofuranInteractswithCalfThymusDNAZHANGLi-jin1,MINShun-geng1,LIGuo-xue2,XIONGYan-mei1,SUNYing1*1.CollegeofScience,ChinaAgriculturalUniversity,Beijing100094,China2.CollegeofResourceandEnvironment,ChinaAgriculturalUniversity,Beijing100094,ChinaAbstractCarbofuranisaninsecticideusedonavarietyofcorps.Acuteandchronicoccupationalexposureofhumanstocarbofuranhasbeenobservedtocausecholinesteraseinhibition,butlittleisknownabouttheinteractionofcarbofuranwithDNA.UsingthetechniqueofUVspectrumandfluorescencequenchingrespectively,theinteractionbetweencarbofuranandctDNAwasstudied.TheUVspectrumshowedthatctDNAcanleadtothehypochromiceffectandredshiftoftheUVspectrumofcarbofuran.Thequenchingprocesswasprovedtobethesinglestaticquenchingandquenchingconstantdecreaseswithtemperatureincreasing.ThebasisofthisspecificityisintercalationofinsecticidebetweenbasepairstoproducectDNA-carbofuranadducts.Furthermore,ethanolcanproduceFranck-con-doneffectonthectDNA-carbofuranadducts.Atdifferentsodiumchlorideconcentrations,quenchingconstanthadnosignificantchange,whichappearedthattherewaslittleelectrostaticinteractionbetweenctDNAandcarbofurananditwasintercalation.KeywordsMechanism;Carbofuran;ctDNAadducts;UVspectrum;Fluorescentquenching;Intercalation(ReceivedNov.18,2003;acceptedMar.6,2004)24725