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ID:33234581
大小:3.80 MB
页数:35页
时间:2019-02-22
《转染survinvin基因的mscs对小鼠肾脏缺血再灌注损伤的影响》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、重庆医科大学硕士学位论文转染Survinvin基因的MSCs对小鼠肾脏缺血再灌注损伤的影响姓名:齐玉增申请学位级别:硕士专业:外科学指导教师:苟欣201206转染Survinvin基因的MSCs对小鼠肾脏缺血再灌注损伤的影响摘要目的:探讨转染survinvin基因后的骨髓间充质干细胞(MSCs)在肾脏缺血微环境中的存活能力以及对小鼠肾脏缺血再灌注损伤的修复作用能力和机制。方法:复苏、扩增培养已完成空病毒、Survinvna基因转染、鉴定、EGFP标记工作的lVlSCs,扩增培养,观察绿色荧光表达情况备
2、用,48只SPF级健康雄性C57BL/6J分为正常对照组12只、瓜组(小鼠双侧肾动脉夹闭40mha后恢复血供)12只、空病毒转染移植组(/VlSCs组,小鼠双侧肾动脉夹闭40min后恢复血供24后经尾静脉注射200微升1×106个细胞)12只、survinvha基因转染移植组(SVV/MSCs组,小鼠双侧肾动脉夹闭40rnin后恢复血供24小时后经尾静脉注射200微升1×106个survinvin基因转染细胞)24只,选取细胞移植后1d、3d、7d、14d不同时间点采集血清行肌酐尿素氮检测,切取小鼠肾
3、脏行石蜡切片观察lVlSCs存活数目并定量分析,HE染色观察肾组织病理变化及肾小管损害程度评分,ELISA检测肾脏匀浆中细胞因子HGF、bFGF、IL一10的表达情况。结果:复苏、扩增培养出的IVISCs增值活力旺盛,荧光状态良好;移植细胞1天后lVISCs组与SVV/IVISCs组肌酐尿素氮水平明显低于R重庆医科大学殒士研究生学位论文对照组(p4、肾组织的移植细胞数目SVV/MSCs组远高于MSCs组,表达EGFP的MSCs主要分布于肾小球周围、小血管内壁、肾小管与肾小管之间的间质,而肾小管内壁几乎见不到表达EGFP的MSCs。在肾脏缺血损伤后保护性因子HGF、bFGF、IL一10水平升高SVV/MSCs也比瓜组、MSCs组明显的多(P5、:骨髓间充质干细胞;缺血再灌注损伤;Survinvin;存活能力EFFECTOFSURVWIN.ENGINEEREDMESENCHYMALSTEMCELLSONRENALISCHEMIA.REPERFUSIONINJURYINMICEABSTRACTObjective:ToinvestigatethesurvivabilityofSurvivin-EngineeredMSCsinmicerenalafterischemia-reperfusioninjuryandexploretherecoverya6、bilityandthepossiblemechanismonrecuperationofthekidneyMethods:ThawingandExpandingMSCs,andMSCsthathadbeeninfectedwiththeseIf-inactivelentiviralvectorGFP—wre—CMLTRandFlap—Ubiqutinpromoter(GCF∽carryingenhancedgreenfluorescentprotein(EGFP)geneandSurvinvinre7、combinantvector(GCFU-svv)ObservetheexpressionofEGFPintwogroupsofMSCsbeforeinjection.Totm48healthymaleC57BL/6JmiceweredividedintofourgroupsNormalcontrolgroups212,IRgroups=12(clampingbilateralrenalpediclesandthenreopeningafter40minutes.Meanwhile,PBSwerein8、jectedthroughcaudalveinintothebodyafteroperation24h),MSCsgroups2:12(clampingbilateralrenalpediclesandthenreopeningafter40minutes.Meanwhile,200al1×106MSCswereinjectedthroughcaudalveinintothebodyafteroperation24h),SVWMSCsgroups=12(
4、肾组织的移植细胞数目SVV/MSCs组远高于MSCs组,表达EGFP的MSCs主要分布于肾小球周围、小血管内壁、肾小管与肾小管之间的间质,而肾小管内壁几乎见不到表达EGFP的MSCs。在肾脏缺血损伤后保护性因子HGF、bFGF、IL一10水平升高SVV/MSCs也比瓜组、MSCs组明显的多(P5、:骨髓间充质干细胞;缺血再灌注损伤;Survinvin;存活能力EFFECTOFSURVWIN.ENGINEEREDMESENCHYMALSTEMCELLSONRENALISCHEMIA.REPERFUSIONINJURYINMICEABSTRACTObjective:ToinvestigatethesurvivabilityofSurvivin-EngineeredMSCsinmicerenalafterischemia-reperfusioninjuryandexploretherecoverya6、bilityandthepossiblemechanismonrecuperationofthekidneyMethods:ThawingandExpandingMSCs,andMSCsthathadbeeninfectedwiththeseIf-inactivelentiviralvectorGFP—wre—CMLTRandFlap—Ubiqutinpromoter(GCF∽carryingenhancedgreenfluorescentprotein(EGFP)geneandSurvinvinre7、combinantvector(GCFU-svv)ObservetheexpressionofEGFPintwogroupsofMSCsbeforeinjection.Totm48healthymaleC57BL/6JmiceweredividedintofourgroupsNormalcontrolgroups212,IRgroups=12(clampingbilateralrenalpediclesandthenreopeningafter40minutes.Meanwhile,PBSwerein8、jectedthroughcaudalveinintothebodyafteroperation24h),MSCsgroups2:12(clampingbilateralrenalpediclesandthenreopeningafter40minutes.Meanwhile,200al1×106MSCswereinjectedthroughcaudalveinintothebodyafteroperation24h),SVWMSCsgroups=12(
5、:骨髓间充质干细胞;缺血再灌注损伤;Survinvin;存活能力EFFECTOFSURVWIN.ENGINEEREDMESENCHYMALSTEMCELLSONRENALISCHEMIA.REPERFUSIONINJURYINMICEABSTRACTObjective:ToinvestigatethesurvivabilityofSurvivin-EngineeredMSCsinmicerenalafterischemia-reperfusioninjuryandexploretherecoverya
6、bilityandthepossiblemechanismonrecuperationofthekidneyMethods:ThawingandExpandingMSCs,andMSCsthathadbeeninfectedwiththeseIf-inactivelentiviralvectorGFP—wre—CMLTRandFlap—Ubiqutinpromoter(GCF∽carryingenhancedgreenfluorescentprotein(EGFP)geneandSurvinvinre
7、combinantvector(GCFU-svv)ObservetheexpressionofEGFPintwogroupsofMSCsbeforeinjection.Totm48healthymaleC57BL/6JmiceweredividedintofourgroupsNormalcontrolgroups212,IRgroups=12(clampingbilateralrenalpediclesandthenreopeningafter40minutes.Meanwhile,PBSwerein
8、jectedthroughcaudalveinintothebodyafteroperation24h),MSCsgroups2:12(clampingbilateralrenalpediclesandthenreopeningafter40minutes.Meanwhile,200al1×106MSCswereinjectedthroughcaudalveinintothebodyafteroperation24h),SVWMSCsgroups=12(
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