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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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低温缺氧条件下脂质体介导的寡核苷酸对人脐静脉内皮细胞-304的转染
, PP. 497-500
Keywords: 脐静脉内皮细胞,低温缺氧保存,脂质体,寡核苷酸,转染效率
Abstract:
目的研究在euro-collins溶液(ecs)中,低温(4℃)缺氧条件下体内阳离子脂质体转染剂(invivoliposome)介导nf-κb诱捕物寡核苷酸(nf-κbdecoyodn)在人脐静脉内皮细胞株ecv-304中的转染效率。方法(1)将ecv-304在含10%胎牛血清的rpmi1640培养液中以37℃、5%co2的条件培养,待生长至单层融合后,进行试验。(2)使用前配制脂质体-odn复合物,脂质体与odn的+/-电荷比率为2。(3)应用荧光显微镜和流式细胞仪,观察在ecs中、4℃条件下,odn浓度分别为0.50、0.75、1.00、1.25μmol/l时,缺氧保存2、4、6、8h后ecv-304的平均荧光强度(mfi)和odn的胞内分布;同时设立裸odn转染为对照组,观察对ecv-304的转染效率。结果在ecs中、4℃缺氧保存条件下,随着保存时间的延长和脂质体-odn浓度的升高,ecv-304的mfi增强,保存6h后mfi基本达到了最大强度,并且大部分odn均位于细胞核内;而odn的细胞摄取率无差异。但与裸转染相比,mfi和细胞摄取率均有显著差异性。结论在ecs中和低温缺氧条件下,脂质体可以高效地将nf-κbdecoyodn转染到ecv-304细胞核,为供体器官在基因调控水平的保护研究提供了实验依据。
| [1] | zabnerj,fasbenderaj,moningert,etal.cellularandmolecularbarrierstogenetransferbyacationiclipid[j].jbiolchem,1995,270(54):18997-9007.
|
| [2] | noguchia,furunot,kawaurac,etal.membranefusionplaysanimportantroleingenetransfectionmediatedbycationicliposomes[j].febslett,1999,433(2):169-73.
|
| [3] | 胡英,金一,王华,等.阳离子膜融合脂质体介导反义寡核苷酸在hela细胞中的转染试验研究[j].药学学报,2002,37(11):892-6.huy,jiny,wangh,etal.studyonthetransfectionefficiencyofantisenseoligodeoxynucleotideagainsttelomerasebycationicfusogenicliposome[j].actapharmsin,2002,37(11):892-6.
|
| [4] | nakanishim,mizuguchiah,ashiharak,etal.genetransfervectorsbasedonsendaivirus[j].jcontrolrel,1998,54(1):61-8.
|
| [5] | harverypr,ius,mckeowncm,etal.adeninenucleotidetissueconcentrationsandliverallograftviabilityaftercoldpreservationandwarmischemia[j].transplantation,1988,45(6):1016-20.
|
| [6] | gibbonsgh,dzauvj.moleculartherapiesforvasculardisease[j].science,1996,272(5262):689-93.
|
| [7] | 郑刚,张积仁,刘发全,等.脂质体介导al,4-galt反义寡核苷酸对人脑胶质瘤细胞系swo-38的作用[j].第一军医大学学报,2002,22(4):299-302.zhengg,zhangjr,liufq,etal.effectsoflipofectin-mediateda1,4galtantisenseoligonucleotideonhumangliomacelllineswo-38[j].jfirstmilmeduniv/diyijunyidaxuexuebao,2002,22(4):299-302.
|
| [8] | 陈永鹏,章廉,胡俊,等.脂质体的乳糖化修饰及其肝脏靶向性研究[j].第一军医大学学报,2001,21(3):177-9.chenyp,zhangl,huj,etal.studyoflactosaminatedliposomeanditshepatotropictargetinginrats[j].jfirstmilmeduniv/diyijunyidaxuexuebao,2001,21(3):177-9.
|
| [9] | 胡俊,章廉,冯筱榕,等.一种新型脂质体的制作方法[j].第一军医大学学报,2001,21(2):138-9huj,zhangl,fengxr,etal.amethodforproducinganewtypeofliposome[j].jfirstmilmeduniv/diyijunyidaxuexuebao,2001,21(2):138-9.
|
| [10] | immaie.genetherapyapproachinrenaldiseaseinthe21stcentury[j].nephroldialtransplant,2001,16(suppl5):26-34.
|
| [11] | liuf,qih,huangl,etal.factorscontrollingtheefficiencyofcationiclipid-mediatedtransfectioninvivoviaintravenousadministration[j].genether,1997,4(5):517-23.
|
| [12] | imaie,isakay.genetherapyforkidneydisease[j].expertopininvestdrugs,2000,9(10):1029-39.
|
| [13] | morishitar,sugimotot,aokim,etal.invivotransfectionofciselement"decoy"againstnuclearfactor-kappabbindingsitepreventmyocardialinfarction[j].natmed,1997,3(8):894-9.
|
| [14] | ingridhcv,rolandg,hermannjg,etal.nf-κbdecoyoligodeoxynucleotidesreducemonocyteinfiltrationinrenalallografts[j].fasebj,2000,14(7):815-22
|
| [15] | barnespj,karinm.nuclearfactor-κb-apivotaltranscriptionfactorinchronicinflammatorydisease[j].newengjmed,1997,336:1066-71.
|
| [16] | collinst.endothelialnuclearfactor-kappabandtheinitiationoftheatheroscleroticlesion[j].labinvest,1993,68(4):499-508.
|
| [17] | cinesdb,pollakes,buckca,etal.endothelialcellsinphysiologyandinthepathophysiologyofvasculardisorders[j].blood,1998,91(363):3527-61.
|
| [18] | tomitan,morishitar,higakij,etal.anovelstrategyforgenetherapyandgeneregulationanalysisusingtranscriptionfactordecoyoligonucleotides[j].expnephrol,1997,5(4):429-34..
|
| [19] | azumah,tomitan,kanedah,etal.transfectionofnfκb-decoyoligodeoxynucleotidesusingefficientultrasound-mediatedgenetransferintodonorkidneysprolongedsurvivalofratrenalallografts[j].genether,2003,10(4):415-25.
|
| [20] | tomitan,morishitar,tomitas,etal.transcriptionfactordecoyfornfkappabinhibitstnf-alpha-inducedcytokineandadhesionmoleculeexpressioninvivo[j].genether,2000,7(13):1326-32.
|
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