补阳还五汤联合骨髓间充质干细胞移植对大鼠脑缺血再灌注损伤的保护作用

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1、摘要ThestudyofNeuroprotectionofBuyanghuanWUsoupAssociatedwithMesemchymalStemCellsTransplantationfocalcerebralischemia／reperfusioninratsMeiXuerui(InternalMedicineofTraditionalChineseMedicine)DirectedbyprofessorZhengShaozhouAbstractObjective：Themodelofacutereperfusioninjuryafterc

2、erebralischemiainratswasestablishedbyunilateralmiddlecerebralartery(MCA)occlusionandbilateralcarotidarteries(CCA)occlusion，toinvestigatetheeffectsandtheprotectivemechanismsofSaponinsfromBuyanghuanWUsoupandMSC··derivedneuron··likecellson．cerebralischemia—reperfusion(I／R)injury

3、．Methods：TheratMSCswereisolatedprimarilyfromthefemursandtibiasofthealbinoSprague—DawleyadultratsandmaintainedinDMEMsupplementedwith10％fetalbovineserum(FBS)．After5daysofincubationat37。C，in5％C02，nonadherentcellswereremovedfromtheculturesbyreplacingmidium．Whentheculturesreachedc

4、onfluency,thecellswereliftedbyincubationwith0．25％trypsinfor2or3minutesatroom÷emperature．The+modeloflocalischemia-reperfusioninratswasestablishedwiththesutureoccludedmethodinventedbyZeaLongaandimprovedproperlyinourlab．MaleSDratswererandomlydividedintofourgroups：ratsingroupl(sh

5、am)onlyreceivedanesthesiaandvesselseparation；ratsingroup2receivedischemiareperfusionoperationandNSig；ratsingroup3isthetranspiantgroupofratMSCs；ratsingroup4isjointgroupfromBuyanghuanWUsoupandMSC-derivedneuron-likecells．TheratsMSCsweretransplantatedintotransplantationgroup，thej

6、ointgroupbythetailvein，thesamevolumeofsalinewasinjectedintosham—operatedgroup，modelgroupbythetailveinafter24hreperfusionfollowing2hischemia．Jointgroupwaseadministratedaccordingtothedoeages(15ml／kg)，theshamgroup，modelgroup，thetransplantgroupweretreatedwithgastricperfusionofdis

7、trilledwaterofthesanevolumerespectively．Theratsinthedaysbeforethesurgerybegandelivery，onceaday．Theneurologicaldeficitscoreofratsineachgroupwasevaluatedafterld，4d，7d．1．补阳还五汤联合骨髓问充质干细胞移植对大鼠脑缺血再灌注损伤的保护作用ischemia’reperfusion．Thenthesubiectswerekilled．Theinfarctvolumeweremeasureda

8、ndpathologicalexaminationwasperformedTheapoptosisoftheneuralcellswas