腺病毒介导NBak基因转移对大鼠红核脊髓束的修复作用

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1、第三军医大学博士学位论文腺病毒介导N-Bak基因转移对大鼠红核脊髓束的修复作用姓名：鞠明兵申请学位级别：博士专业：外科学（野战外）指导教师：廖维宏20040501第三军医大学博士学位论文AmpAmpicillin氨苄青霉素KanKanamycin卡那霉素LBLuria-BertaniculturemediumLB培养基DMEMDulbecco’smodifiedessentialmediumDMEM培养基Bcl-2Bcelllymphoma/leukemia-2geneB细胞淋巴瘤/白血病-2基因TUNELTerminaldeoxynucleotidaltransferase-medi

2、ated末端脱氧核苷酸转移酶介导dUTP-biotinnickendlabeling的dUTP缺口末端标记2第三军医大学博士学位论文EffectsofadenovirusN-BakgenetransferonrubrospinaltractinjuryinratsAbstractItisestimatedthattheannualincidenceofspinalcordinjury(SCI)isapproximately50casespermillionpopulationanditisgettingupinrecentyears.SCIprimarilyaffectsyoungad

3、ults.Thecostlyexpensesandhandicapaftertheinjuryhasbeenanoverwhelmingfinancialandsocialburdenonthefamiliesandnations.Thissignalsanurgentneedforstudiesonitsbasicalresearcherandclinicaltreatments,soastoimproverepairandrecoveryafterSCI.ThetreatmentofSCIisbasedonprotectionagainstdelayedneuronaldeath.

4、Inrecentexperimentaltreatment,severalstrategies,suchasneuralnutritionalfactorssupply,genetransfer,tissuefactorreplacement,havebeenappliedtopreventdelayedneuralinjury,tofacilitateaxonregeneration,andtoimprovethemicroenvironmentandinterferegeneexpressioninneurons.Moreover,theaxongrowstotargetcells

5、andreconstructtheanatomicconnectionsarethekeypointsofthefunctionalrestoration,whichisinterferedbyphysicalandchemicalbarrierconstructedbyglialscar.Howtosolvethisproblemhasimportantsignificanceinfunctionalrestoration.N-BakgenewasdiscoveredandrenamedbyArumaeandcolleaguesin2001intheirresearchofBakge

6、neexpressionandfunctioninneurons.EndogeneticN-BakmRNAisexpressedexclusivelyinneurons,andits22kDtranscriptionalproductonlycontainsBH3domain.However,differentfromotherBcl2familymembers,N-BakisantiapoptoticwhenoverexpressedinNGF-deprivedneurons,whereasitefficientlykillstransfectednonneuronalcells(s

7、uchasCOS7andHeLacells).IthasbeendemonstratedbymutationanalysisthatanintactBH3domainissufficienttokillthenonneuronalcells,whereasotherdomainsinadditiontotheBH3domainarenecessarytoprotectapoptoticneurons.Inthisstudy,weconstruc