・246・中华高血压杂志2009年3月第17卷第3期ChinJHypertens,March2009,Vol.17No.3・论著・血管内皮生长因子激活钙调神经磷酸酶促进内皮祖细胞增殖杨龙1,杨新春2,杨军珂3【摘要】背景血管内皮生长因子(VEGF)在内皮祖细胞(EPC)生物活性的调控中涉及多条信号通路,但是否通过调控钙调神经磷酸酶(CaN)信号通路而影响EPC活性尚不清楚。目的探讨钙调神经磷酸酶(CaN)在血管内皮生长因子(VEGF)调节EPC增殖活性中的作用。方法采用密度梯度离心法分离外周血单个核细胞,贴壁培养获得EPC,分四组:对照组;VEGF组(VEGF50μg/L干预时间48h);环孢素A组(CsA10mg/L干预48h);环孢素A+VEGF组(CsA10mg/L干预1h,再予VEGF50μg/L干预时间48h)。比色法测定CaN活性及EPC增殖能力;TUNEL染色检测EPC凋亡;逆转录聚合酶链法检测bcl2和bax基因表达。结果VEGF刺激明显增强CaN活性(与对照组比较增加73%,P<0101),促进EPC增殖(与对照组比较增加89%,P<01),增强bcl2基因表达(与对照组比较增加40%,P<0105)。CsAVEGF组与VEGF组比较,CaN活性减弱60%(P<0101),增殖能力减弱78%(P<0101)101)],并增加细胞凋亡(凋亡率:CsA组:2317%比对照组:13%,)V和bax基因表达无明显影响。结论VEGF可通过激活【关键词】;;钙调神经磷酸酶VascularEndothelialGrowthFactorPromotestheProliferativeCapacityofEndothelialProgenitorCellsbyIncreasingCalcineurinEnzymaticActivityYANGLong1,YANGXin2chun2,YANGJun2ke3.1.Departmentofcardiolo2gy,ZunyiHospital,ZunyiGuizhou563002;2.CenterofCardiology,BeijingChaoyangHospitalofCapitalMedicalUniversity,Beijing100000;3.DepartmentofCardiology,theGeneralHospitalofSecondArtilleryofPLA,Beijing100000;China【Abstract】BackgroundVascularendothelialgrowthfactor(VEGF)hasbeenshowntocross2talkwithnu2meroussignalingpathwaysonregulatingthebiologicalfunctionofendothelialprogenitorcells(EPC).LittleisknownabouttheroleofVEGFonregulatingtheactivationofcalcineurinsignalinEPC.ObjectiveToinvestigatewhethervascularendothelialgrowthfactor(VEGF)regulatestheproliferativecapacityofendothelialprogenitorcells(EPC)bythewayofactivatingcalcineurin(CaN)signal.MethodsEPCwereobtainedfromculturedmononuclearcellsisolatedfromperipheralbloodofhealthyadults,andwerereceivedfollowingapproaches:Control,VEGF(50μg/L),cyclosporinA(CsA,10mg/L),andCsA+VEGF(incubationwithCsAfor1hbeforeVEGFwasadd2ed).CaNenzymaticactivityandcellproliferationwereassayedusingcolorimetricmethod.Cellapoptosiswasde2terminedbyTUNELstaining.Theexpressionofbcl2andbaxgeneswasinvestigatedbyRT2PCRanalysis.ResultsComparedwiththecontrolgroup,VEGF(50μg/L)significantlyincreasedCaNenzymaticactivity(+73%,P<0101),cellproliferation(+89%,P<0101),andbcl2geneexpression(+40%,P<0101).Pre2treatedwithcyclosporine(10mg/L)markedlyabrogatedtheaforementionedeffectsofVEGF.ComparedwiththeVEGFgroup,CsA+VEGFdecreasedtheCaNenzymaticactivity,cellproliferation,andbcl2geneexpression60%,78%and82%,respectively,allP<0101)andincreasedtheapoptoticrateofEPC.Nostatisticsignificantdiffer2enceofbaxgeneexpressionwasfoundbetweengroups.ConclusionVEGFtreatmentpromotesproliferativecapac2ityofhumanEPCbyactivatingCaNsignal.【Keywords】Endothelialprogenitorcells;Vascularendothelialgrowthfactor;Calcineurin收稿日期:2008209229作者单位:11贵州省遵义医院心内科,贵州遵义563002;21首都医科大学附属北京朝阳医院心脏中心,北京100000;31中国人民解放军第二炮兵总医院心内科,北京100000通信作者:杨新春,电话:010285231586;E2mail:yanglong1001@163内皮功能损害是动脉硬化及原发性高血压发生、发展的重要因素。血管内皮祖细胞(EPC)是成熟内皮细胞的前体细胞,在修复损伤血管内皮、维护机体内皮©1994-2009ChinaAcademicJournalElectronicPublishingHouse.Allrightsres...