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时间:2020-05-04
《不同转染途径对慢病毒载体基因在大鼠肝脏中表达的影响-论文.pdf》由会员上传分享,免费在线阅读,更多相关内容在应用文档-天天文库。
1、·96·doi10.3969~.issn.1673—4254.2014.01.21JSouthMedUniv,2014,34(1):96—99基础研究不同转染途径对慢病毒载体基因在大鼠肝脏中表达的影响赵英鹏,李立,马菁番,白建华,刘其雨昆明市第一人民医院暨昆明医科大学附属甘美医院肝胆胰外科,云南昆明650011摘要:目的研究慢病毒载体基因在肝脏内转染的有效途径。方法本实验根据大鼠消化系统解剖,设计利用经回盲部回结肠静脉穿刺置管至门静脉作为转染途径。通过观察对比尾静脉转染途径大鼠肝脏荧光蛋白表达、基因表达及转氨酶变化情况,探讨不同转染方
2、法的有效性和安全性。结果门静脉转染组和尾静脉转染组大鼠均存活。转染后96h慢病毒载体携带绿色荧光蛋白在门静脉转染组肝脏呈高表达,14dR仍有持续表达;尾静脉转染组术后96h荧光蛋白表达较门静脉组弱,14dR几无持续表达。RT-PCR检测转染基因LXRa—RNAi效率:转染组LXRamRNA表达均有下调,门静脉转染组LXRo.~因敲减率明显高于尾静脉转染组(0.135~0.0020.713~0.036,P3、的情况下提高基因转染率,且对肝功能无明显损害,是值得推广的方法。关键词:慢病毒载体;转染;肝脏EffectofdifferentapproachesoflentiviralvectortransfectionontargetgeneexpressioninratliverZHAOYingpeng,LILi,MAlingpan,BAIJianhua,LIUQiyuDepartmentofHepatobiliaryandTransplantationSurgery,GanmeiHospitalAfiliatedtoKunmingMedic4、alUniversity,Kunming650011,ChinaAbstract:0bjectiveToinvestigatetheoptimalapproachof1entiviralvectortransfectionforeffectivedeliveryofexogenousgeneintotheliver.MethodsThe1entiviralvectorwasdeliveredviatheileocolicveinoftheileocecus(portalveingroup)orviathecaudalveinofSDr5、ats.Theeffectgenetransfectionintotheliverwasassessedbyobservingtheexpressionofgreenfluorescenceproteinexpressioncarriedbythelentiviralvector,silencingofLXRamRNAexpressionmediatedbyRNAinterference,andlivertransaminasechanges.Theefficiencyandsafetvofthetwoapproachesoftran6、sfectionwereevaluated.ResultsA11theratsreceivinglentiviraltransfectionsurvived.Intheportalveingroup,abundantgreenfluorescencewasdetectedintheliverat96hfollowingthetrasnfectionandlastedtill14days,whereasonlyweakfluorescencewasobservedinthecaudalveingroup.TheresultsofRT—P7、CRdemonstratedasignificanthigherrateofLXRknock—downinportalveingroupthanincaudalveingroup(0.135~0.002VS0.713~0.036,P<0.05).NOsignificantdifferenceinALTlevelsfoundbetweenthetwogroups.ConclusionInfusionviathepota1veiniseffectiveforgenetransfectionintotheliver,andpuncturef8、romtheileocolicveinofileocecuscanguaranteethesurvivalofratsandimprovethetransfectionefficiencywithoutcausingli
3、的情况下提高基因转染率,且对肝功能无明显损害,是值得推广的方法。关键词:慢病毒载体;转染;肝脏EffectofdifferentapproachesoflentiviralvectortransfectionontargetgeneexpressioninratliverZHAOYingpeng,LILi,MAlingpan,BAIJianhua,LIUQiyuDepartmentofHepatobiliaryandTransplantationSurgery,GanmeiHospitalAfiliatedtoKunmingMedic
4、alUniversity,Kunming650011,ChinaAbstract:0bjectiveToinvestigatetheoptimalapproachof1entiviralvectortransfectionforeffectivedeliveryofexogenousgeneintotheliver.MethodsThe1entiviralvectorwasdeliveredviatheileocolicveinoftheileocecus(portalveingroup)orviathecaudalveinofSDr
5、ats.Theeffectgenetransfectionintotheliverwasassessedbyobservingtheexpressionofgreenfluorescenceproteinexpressioncarriedbythelentiviralvector,silencingofLXRamRNAexpressionmediatedbyRNAinterference,andlivertransaminasechanges.Theefficiencyandsafetvofthetwoapproachesoftran
6、sfectionwereevaluated.ResultsA11theratsreceivinglentiviraltransfectionsurvived.Intheportalveingroup,abundantgreenfluorescencewasdetectedintheliverat96hfollowingthetrasnfectionandlastedtill14days,whereasonlyweakfluorescencewasobservedinthecaudalveingroup.TheresultsofRT—P
7、CRdemonstratedasignificanthigherrateofLXRknock—downinportalveingroupthanincaudalveingroup(0.135~0.002VS0.713~0.036,P<0.05).NOsignificantdifferenceinALTlevelsfoundbetweenthetwogroups.ConclusionInfusionviathepota1veiniseffectiveforgenetransfectionintotheliver,andpuncturef
8、romtheileocolicveinofileocecuscanguaranteethesurvivalofratsandimprovethetransfectionefficiencywithoutcausingli
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