异体骨髓干细胞移植后假肥大型肌营养不良症模型鼠膈肌dystrophin表达及病理改变作者：张雅妮；张成；于美娟；王淑辉；李美山；黄慧；熊符；冯善伟；柳太云；卢锡林(中山大学第一医院神经科，广东广州510080)摘要：目的探讨骨髓干细胞移植对假肥大型肌营养不良症(DMD)模型鼠-mdx鼠膈肌的治疗效果。方法取雄性SD大鼠骨髓干细胞经尾静脉植入放疗处理后的8周龄雌性mdx鼠(n=18)，于移植后4、8、12周各取6只mdx鼠的膈肌行HE染色、抗肌萎缩蛋白(dystrophin)免疫荧光检测以及dystrophinmRNA的RT-PCR分析，同时用正常C57鼠及放疗而未移植的mdx鼠作为对照，另进行PCR反应检测实验鼠膈肌内Sry(Y染色体的性别决定区)基因。结果移植后mdx鼠膈肌间质内炎性细胞浸润较放疗而未移植mdx鼠有所减少；移植后核中心移位纤维比例[移植后4、8、12周分别为(15.58±0.91)%、(12.50±1.87)%、(10.17%±1.17)%]较未移植mdx鼠(19.5±1.87)%显著减少。移植后dystrophin免疫荧光阳性细胞比例[移植后4、8、12周分别为(1.00±0.32)%、(6.00±1.05)%、(11.92±1.11%)]较未移植mdx鼠(0.17±0.41)%显著增加。RT-PCR结果显示C57鼠膈肌的dystrophinmRNA相对含量(0.63±0.04)最高；而未移植mdx鼠的膈肌中未检测到dystrophinmRNA；移植后的mdx鼠膈肌中mRNA的表达水平(移植后4、8、12周分别为0.19±0.05、0.26±0.06、0.36±0.04)随时间推移逐渐增高；移植后各时间点mdx鼠膈肌Sry基因均为阳性。结论骨髓干细胞系统移植mdx鼠可以恢复部分膈肌的dystrophin表达，改善膈肌的病理,骨髓干细胞移植有希望成为全身治疗DMD的有效方法。关键词：假肥大型肌营养不良症；mdx鼠；膈肌；骨髓移植；抗肌萎缩蛋白：R392.4;R457.7文献标识码：A：1673-4254(2006)01-0053-06DystrophinexpressionandpathologyofdiaphragmmusclesofmdxmiceafterxenogenicbonemarrowstemcelltransplantationZHANGYa-ni；ZHANGCheng；YUMei-juan；WANGShu-hui；LIMei-shan；HUANGHui；XIONGFu；FENGShan-wei；LIUTai-yun；LUXi-linDepartmentofNeurology,FirstHospitalofSunYat-senUniversity,Guangzhou510080,ChinaAbstract:ObjectiveToinvestigatetheeffectofbonemarrowstemcelltransplantation(BMT)onthediaphragmmusclesofmdxmice,amousemodelofDuchennemusculardystrophy(DMD).MethodsThebonemarrow-derivedstemcellsformmaleSDratswastransplantedthroughthetailveininto18female8-week-oldmdxmice,whichweresacrificedat4,8and12weeksafterBMT(6ateachtimepoint),respectively.ThediaphragmmusclesofthemiceweresubjectedtoHEstaining,immunofluorescencedetectionofdystrophin,reversetranscription(RT)-PCRanalysisofdystrophinmRNAtranscriptsandPCRanalysisofSry(sex-determiningregionontheYchromosome)gene,withage-matchedfemaleC57miceanduntreatedmdxmiceasthecontrols.ResultsTheproportionofcentrallynucleatedfibers(CNF)inthediaphragmmuscleoftherecipientmdxmicewas(15.58±0.91)%,(12.50±1.87)%and(10.17±1.17)%at4,8and12weeksafterBMT,respectively,significantlysmallerthanthatofuntreatedmdxmice[(19.5±1.87)%],andthefibersafterBMTshowedlessinflammatoryinfiltration.Comparedwiththeuntreatedmice,therecipientmdxmiceshowedgreenfluorescenceonsignificantlymorediaphragmmusclecellmembranes[withtheproportionofdystrophin-positivefibersof(1.00±0.32)%,(6.00±1.05)%and(11.92±1.11)%at4,8,and12weeksafterBMT].RT-PCRofdystrophinmRNAalsodemonstratedsignificantlyhigherrelativelevelsofdystrophinintherecipientmdxmice(0.19±0.05,0.26±0.06and0.36±0.04at4,8and12weeksafterBMT)thaninuntreatedmdxmice,andSrygenewaspresentintherecipientmice.ConclusionsBMTcanpartiallyrestoredystrophinexpressionandamelioratethepathologyinthediaphragmmusclesofmdxmice,andhasgreatpotentialtoproducegeneraltherapeuticeffectinpatientswithDMD.Keywords:Duchennemusculardystrophy;mdxmice;diaphragm;bonemarrowtr...