降钙素基因相关肽调节间充质干细胞存活对大鼠颈动脉损伤后血管修复的作用研究赵然尊，朱江兰，王冬梅，刘志江，盛瑾，石蓿,（遵义医学院附属医院心血管内科，贵州遵义563003）［摘要］目的探讨降钙素基因相关肽（CalcitoninGene-relatedPeptide,CGRP）调节间充质干细胞（MesenchymalStemCells,MSCs）生存和对大鼠颈动脉损伤后血管修复作用及其可能机制。方法分离培养获得大鼠MSCs,分为CGRP组和对照组，均给予过氧化氢（H2O2）处理后，前者再同时加入CGRP处理，以流式细胞仪检测MSC的存活和凋亡情况，并以Westernblot检测磷酸化GSK3/3（pGSK3/3）和/^catenin的表达情况。构建大鼠颈动脉球囊损伤模型，分为MSCs组和对照组，分别给予MSC-CGRP（以携带CGRP的重组慢病毒载体pLen-CGRP-EGFP转染细胞）和MSC（未携带CGRP的重组慢病毒载体pLen-EGFP转染细胞）移植治疗，以Westernblot检测血管局部CGRP的表达以及pGSK3/3和/3-catenin的表达情况；免疫荧光染色检测损伤血管CD31的表达；HE染色检测损伤血管新生内膜/中膜面积。结果MSCs给予H2O2处理后显著促进细胞凋亡；而H2O2处理的MSC-CGRP则细胞凋亡明显减少，伴随着pGSK3/5和/9catenin表达增加。在大鼠颈动脉损伤模型中，MSCs移植可促进血管内皮修复，改善动脉重构；而MSC-CGRP移植较MSCs应用后损伤动脉的内皮化程度更高，伴随着局部PGSK3/5和/^catenin蛋白表达增加。结论CGRP可能通过激活GSK3/3/^catenin信号通路，促进移植的MSCs存活并增强其对动脉损伤后血管修复作用，这为防治血管增殖性疾病提供治疗思路。［关键词］降钙素基因相关肽；间充质干细胞；血管修复；细胞存活［中图文分类号］R543.4［文献标识码］A［基金来源］国家自然科学基金（NSFC30860100）；贵州省国际合作项目（黔科合外G字［2010］0732）ThesurvivaleffectofCalcitoninGene-relatedPeptidetoMSCanditsvascularrepairintheratsofcarotidarteryinjurymodelZhaoRanzun,ZhuJianglan,WangDongmei,LiuZhijiang,Shengjin,ShiBei(DepartmentofCardiology,theAffiliatedHospitalofZunyiMedicalUniversity,ZunyiGuizhouProvince,563000)[Abstract]ObjectiveToinvestigatetheeffectofCalcitoninGene-relatedPeptide(CGRP)tothesurvivalofMesenchymalstemcells(MSCs)anditsvascularrepairaftercarotidarteryinjuryinratsanditspossiblemechanism.MethodsMSCsweredividedintoCGRPgroupandcontrolgroup,andthecellsweretreatedwithH2O2inbothgroupsandthenweretreatedwithCGRPintheCGRPgroup.ThesurvivalandapoptosisofMSCswereanalyzedbyflowcytometry,andthenn±i矗redetectedbyWesternblot.TheratswereinducedintocarotidarteryinjurymodelanddividedintoMSCgroupandcontrolgroup,treatedwithpLen-CGRP-EGFPorpLen-EGFP,respectively.TheproteinexpressionofCGRP,pGSK3/9and/3-cateninintheinjuredcarotidweremeasuredbywesternblot,theCD31byimmunofluorescencestainingandthearearatioofneointima/tunicamediabyHEstaining.ResultsMSCssignificantlywereinducedcellapoptosis,whereastheapoptosisofMSC-CGRPdecreasedsignificantlyafterH2O2treatment,accompaniedbytheincreaseoftheproteinexpressionofpGSK3ySand&catenin.Intheratsofcarotidarteryinjurymodel,MSCstransplantationcanpromotevascularendothelialrepair,improvearterialreconstruction.There-endotheliulizationintheinjuredarterywashigherintheMSC-CGRPgroupthanthatinMSCgroup,accompaniedbytheincreaseoftheproteinexpressionofpGSK3/9and0catenin.ConclusionCGRPmaybepromotethetransplantationofMSCssurvivalandenhanceitseffectonvascularrepairafterarterialinjurythroughtheactivationofGSK3B/-c建eninsignalingpathway,whichprovidesusthenewwaysinthepreventionandtreatmentofvascularhyperplasticdiseases・[Keywords]CalcitoninGene-relatedPeptide(CGRP);Mesenchymalstemcells(MSCs);Vascularrepair;cellsurvival动脉内皮损伤伴随内皮功能丧失是血管增殖性疾病重要发病机制，如动脉粥样硬化...