用于防齲DNA疫苗粘膜給藥的陽離子脂質(zhì)體載體的研究.pdf

1、華中科技大學(xué)碩士學(xué)位論文用于防齲DNA疫苗粘膜給藥的陽離子脂質(zhì)體載體的研究姓名：奚穎霞申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：生化與分子生物學(xué)指導(dǎo)教師：甘璐20090518II華中科技大學(xué)碩士學(xué)位論文AbstractItisakeyfactfsuccessfulgenetherapytodevelopefficientsafegenedeliverycarriersthatcantransferDNAintothenucleiofthetargetce

2、lls.Cationicliposomeisoneofthemostpromisingnonviralsystemsfgenetherapy.CationicliposomeDNAcomplexes，alsoknownaslipoplexes，areselfassemblingnanosystems.TheycantransferDNAintoalargevarietytypesofcells.Inthepresentstudywepr

3、eparedDOTAPDOPEliposomesacterizedtheinteractionsbetweenthesecationicliposomesplasDNAbyphysicochemicalresearchesstudiedtheirtransfectionefficiencyinvitro.Themainresultsareasfollows:(1)DOTAPDOPEcationicliposomeswereprepare

4、dbythemethodoffilmdispersioninadditionofsonication.Weinvestigatedthetimeofhydrationsonicationofthepreparationtechnology.Severaldatawereacterizedsuchasparticlesizezetapotentialdistributionmphologypreliminaryphysicalstabil

5、ity.Itwasfoundthattherepeatabilityofthepreparationtechnologywaswell.Theresultingsuspensionhadparticlesizeof128.64.6nmPDIof0.1680.016zetapotentialof68.73.8mV.afterthestageat4℃37℃fonemonththecolloidwasstabile.(2)Cationicli

6、posomeDNAcomplexeswerepreparedtheirphysicochemicalpropertieswereacterized.ItwasshownthatDNAfmsstablepositivelygedcomplexeswithcationicliposomesatNPratiosabove1.5byagarosegelelectrophesisparticlesizezetapotentialmeasureme

7、nts.NucleasedigestionassayshowedthatthecomplexesprotectedDNAfromenzymaticdigestion.wealsoinvestigatedtheparticlesizezetapotentialofthelipoplexesthatpreparedatphysiologicalionstrength.ThecomplexeswerefoundpositiveatNPrati

8、osabove3aggregatedtofmlargesizeparticles.Inadditionwepreparedlipoplexeswhichrequiredftheinvivoresearchoftheanimalexperiment.(3)MTTassaytransfectionstudiesinCHONIH3T3cellsshowedthatthecytotoxicityofthecomplexeswererelativ