Sonic Hedgehog Gene Delivery to the Rodent Heart Promotes Angiogenesis via iNOSNetrin-1PKC Pathway 英文参考文献.docVIP

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Sonic Hedgehog Gene Delivery to the Rodent Heart Promotes Angiogenesis via iNOSNetrin-1PKC Pathway 英文参考文献.doc

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Sonic Hedgehog Gene Delivery to the Rodent Heart Promotes Angiogenesis via iNOSNetrin-1PKC Pathway 英文参考文献

SonicHedgehogGeneDeliverytotheRodentHeart PromotesAngiogenesisviaiNOS/Netrin-1/PKCPathway RafeeqP.H.Ahmed,KhawajaHusnainHaider,JiangShujia,MuhammadRizwanAfzal,Muhammad Ashraf* DepartmentofPathologyandLaboratoryMedicine,UniversityofCincinnati,Cincinnati,Ohio,UnitedStatesofAmerica Abstract Background: We hypothesized that genetic modification of mesenchymal stem cells (MSCs) with Sonic Hedgehog (Shh) transgene, a morphogen during embryonic development and embryonic and adult stem cell growth, improved their survivalandangiogenicpotentialintheischemicheartviaiNOS/netrin/PKCpathway. Methods/PrincipalFindings:MSCsfromyoungFisher-344ratbonemarrowwerepurifiedandtransfectedwithpCMVShh plasmid(ShhMSCs).Immunofluorescence,RT-PCRandWesternblottingshowedhigherexpressionofShhin ShhMSCswhich alsoledtoincreasedexpressionofangiogenicandpro-survivalgrowthfactorsin ShhMSCs.Significantlyimprovedmigration and tube formation was seen in ShhMSCs as compared to empty vector transfected MSCs (EmpMSCs). Significant upregulationofnetrin-1andiNOSwasobservedin ShhMSCsinPI3KindependentbutPKCdependentmanner.Forinvivo studies,acutemyocardial infarction modelwasdevelopedin Fisher-344rats. Theanimalsweregroupedtoreceive70 ml basal DMEM without cells (group-1) or containing 16106 EmpMSCs (group-2) and Shh MSCs (group-3). Group-4 received recombinantnetrin-1proteininjectionintotheinfarctedheart.FISHandsry-quantificationrevealedimprovedsurvivalof ShhMSCs post engraftment. Histological studies combined with fluorescent microspheres showed increased density of functionally competent blood vessels in group-3 and group-4. Echocardiography showed significantly preserved heart functionindicespostengraftmentwith ShhMSCsingroup-3animals. Conclusions/Significance:ReprogrammingofstemcellswithShhmaximizestheirsurvivalandangiogenicpotentialinthe heartviaiNOS/netrin-1/PKCsignaling. Citation:AhmedRPH,HaiderKH,ShujiaJ,AfzalMR,AshrafM(2010)SonicHedgehogGeneDeliverytotheRodentHeartPromotes