人血管内皮细胞生长因子-165和人血管生成素-1促进大鼠缺血下肢血管新生Vascular endothelial grow factor 165 and angiopoitin-1 augment revascularization in a rat hindlimb ischemia model
宋建蓉,杨志健,张馥敏,马文珠,哈团柱,李传富,高翔
摘要(Abstract)：
目的 观察肌肉转染腺病毒(adenovirus ,Ad )携带的人血管内皮细胞生长因子- 1 6 5(ves cularendothelialgrowfactor ,Ad5 VEGF1 65)和人血管形成素- 1 (angiopoietin- 1 ,Ad5 Ang -1 )基因对大鼠缺血下肢的生血管效应。方法 制作大鼠下肢血管闭塞模型,肌肉内注射Ad5 VEGF1 65 或 和Ad5 Ang -1 ,以Westernblot法检测生长因子蛋白表达,免疫组化检测基因导入后在缺血肌肉引起的效应。结果 肌肉经注射Ad5 VEGF1 65和Ad5 Ang- 1后高表达人VEGF1 65 蛋白和人Ang- 1蛋白。术后7d处理组与对照组新生毛细血管数目无明显差别。但术后1 4d和2 1dAng- 1和VEGF1 65 组的新生毛细血管 肌肉数目比明显高于对照组(P <0 . 0 1 ) ,VEGF1 65+Ang 1两因子合用组明显高于单用组(P <0. 0 1 )。Ad -VEGF1 65组及Ad VEGF1 65+Ad Ang 1合用组出现大量包围着一厚层αSMA阳性平滑肌细胞的血管,其数量与肌肉数比值明显高于对照组(P <0 . 0 1 )。因子单用组和合用组肌肉内出现大量溴化脱氧尿嘧啶阳性细胞浸润,部分细胞表面呈现C- Kit阳性,新生血管内皮细胞中也有C Kit阳性标记。结论 Ad5- VEGF1 65和Ad5 -Ang -1能促进缺血肢体血管新生,二者合用的生血管效应更为显著;VEGF1 65能促进包被平滑肌细胞的形似微小动脉的血管数量增多
关键词(KeyWords)： 血管内皮细胞生长因子;血管形成素-1;血管生成;基因疗法
基金项目(Foundation): 江苏省科委自然科学基金(BK2001118);; 江苏省卫生重点项目(H2004)
作者(Author): 宋建蓉,杨志健,张馥敏,马文珠,哈团柱,李传富,高翔
参考文献(References)：
- 1ShimpoM ,IkedaU ,MaedaY ,etal.AAV mediatedVEGFgenetransferintoskeletalmusclestimulatesangiogenesisandimprovesbloodflowinarathindlimbischemiamodel.CardiovascularRes,2002,53:9931001.
- 2BaumgartnerI ,PiezekA ,MaorO ,etal.ConstitutiveexpressionofphVEGF165 afterintramusculargenetransferpromotescollateralvesseldevelopmentinpatientwithcriticallimbischemia.Circulation,1998,97:11141123.
- 3ShyuKG ,ManorO ,MagnerM ,etal.DirectintramuscularinjectionofplasmidDNAencodingangiopoietin1,butnotangiopoietin2geneaugmentsrevascularizationinrabbitischemichindlimbmodel.Circulation,1998,98:20812087.
- 4SuriC ,McClainJ ,ThursyonG ,etal.Increasedvascularizationinmiceoverexpressingangiopoietin1.Science,1998,282:468471.
- 5LosordoDW ,VslePR ,SymesJF ,etal.GenetherapyformyocardialangiogenesisinitialclinicalresultswithdirectmyocardialinfarctionofphVEGF A165assolotherapyformyocardialischemia.Circulation,1998,98:28002804.
- 6SuH ,LuR ,KanYW .Adeno associatedviralvector mediatedvascularendothelialgrowthfactorgenetransferinducesneovascularformationinischemicheart.ProcNatlAcadSciUSA ,2000,97:1380113806.
- 7ArsicN ,ZentilinL ,ZacchignaS ,etal.InductionoffunctionalneovascularizationbycombinedVEGFandangiopoietin1genetransferusingAAVvectors.MolTher,2003,7:450459.
- 8LiTS ,HamanoK ,NishidaM ,etal.CD117+stemcellplayakeyroleintherapeuticangiogenesisincludedbybonemarrowdellimplantation.AmJphysiologyHeartCircPhysiol,2003,285:H931H937.
- 9MooreMA ,HattoreK ,HeissigB ,etal.MobilizationofendothelialandhematopoieticstemandprogenitorcellsbyAdenovector mediatedelevationofserumlevelsofSDF 1,VEGF ,andAngiopoietin1.AnnAcadSci,2001,938:3647.
- 10NykanenAI,KrebsR ,SaaristoA ,etal.Angiopoiein1protectsagainstthedevelopmentofcardiacallograftarteriosclerosis.Circulation,2003,10:13081314.
- 11ZhouL ,ZhangF ,LuL ,etal.Cloningandconstructionofadenovirusexpressinghumanangiopoietin1orvascularendotheliaogrowthfactor.ChinJClinRehabil,2003,7:434436.
- 12GoodellMA ,BroseK ,ParadisG ,etal.Isolationandfunctionalpropertiesofmurinehematopoieticstemcellsthatarereplicatinginvivo.JExpMed,1996,183:17971806.
- 13ChaeJK ,KimI ,LimST ,etal.Coadministrationofangiopoietin1andvascularendothelialgrowthfactorenhancescollateralvascularizationarteriosclerosisthrombosisandvascular.Biology,2000,20,25732578.
- 14ZhuWH ,MaclntyreA ,NicosiaRF .Regulationofangiogenesisbyvascularendothelialgrowthfactorandangiopoietin1intherataortamodel.AmJPathol,2002,161:823830.
- 15ShyuKG ,ChangH ,WangBW ,etal.Intramuscularadministrationofcytokinesonrevascularizationoftheischemichindlimbmodelofhyperlipidemicrabbitisfeasible.LifeScience,2003,73:563579.
- 16PuriMC ,RossantJ ,AlitaloK ,etal.ThereceptortyrosinekinaseTIEisrequiredforintegrityandsurvivalofvascularendothelialcells.EMBOJ ,1995,14,58345891.
- 17KranzA ,MayrU ,FrankH ,etal.Thecoronaryendothelium:atargetforvascularendothelialgrowthfactor.Humancoronaryarteryendothelialcellsexpressfunctionalreceptorsforvascularendothelialgrowthfactorinvitroandinvivo.LabInvest,1999,79:985991.
- 18ErikssonU ,AlitaloK .VEGFreceptor1stimulatesstem cellrecruitmentandnewhopeforangiogenesistherapies.NatMed,2002,8:775777.
- 19HattoriK ,DiasS ,HeissigB ,etal.Vascularendothelialgrowthfactorandangiopoietin1stimulatepostnatalhematopoiesisbyrecruitmentofvasculogenicandhematopoieticstemcells.JExpMed,2001,193:10051014.
- 20GrosskreutzCL ,AnandApteB ,DuplaaC ,etal.Vascularendothelialgrowthfactor inducedmigrationofvascularsmoothmusclecellsinvitro.MicrovascRes,1999,58:128136.
- 21IshidaA ,MurrayJ ,SaitoY ,etal.Expressionofvascularendothelialgrowthfactorreceptorsinsmoothmusclecells.JCellPhysiol,2001,188:359368.
- 22ParentiA ,BrogelliL ,FilippiS ,etal.EffectofhypoxiaandendotheliallossonvascularsmoothmusclecellresponsivenesstoVEGF A :roleofflt1VEGF receptor1.CardiovascRes,2002,55:201212.
- 23CucinaA ,BorrelliV ,RandoneB ,etal.Vascularendothelialgrowthfactorincreasesthemigrationandproliferationofsmoothmusclecellsthroughthemediationofgrowthfactorsreleasedbyendothelialcells.JSurgRes,2003,109:1623.
- 24SpringerML ,OzawaCR ,BanfiA ,etal.LocalizedarterioleformationdirectlyadjacenttothesiteofVEGF Inducedangiogenesisinmuscle.MolTher,2003,7:441449.