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1、EphB6在小鼠骨髓间充质干细胞成骨分化中的调控作用李松涛1,刘涌1,宋磊1,董世武2,邸宁1,李殿威1,徐源1,周强1(400038重庆,第三军医大学西南医院骨科,全军矫形外科中心)[摘要]目的本研究旨在了解EphB6在小鼠骨髓间充质干细胞成骨分化中的表达变化及其对成骨分化的影响。方法将小鼠骨髓间充质干细胞通过双相接种法接种于脱钙骨基质构建组织工程骨,分为4组:成骨诱导5d(A)、14d(B),普通培养5d(D)、14d(D),检测各组碱性磷酸酶活性,Runx2、Osterix和EphB6的m
2、RNA表达,然后用游离态配体ephrinB1-Fc激活EphB6并重复检测上述指标。结果ALP活性、Runx2和Osterix在A、B组增高,且B组检测到最高的ALP活性[(7.32±2.02)金氏单位/100ml、Runx2(3.4426±0.2585)倍和Osterix(2.4823±0.3292)倍,EphB6的表达水平在A、B组降低,且在B组最低(0.7543±0.0235)倍。以ephrinB1-Fc激活EphB6后,A、B组EphB6的表达较试剂空白上调(P<0.05),ALP活性显
3、著下降(P<0.05),在高浓度配体作用后ALP活性为(12.20±1.30)金氏单位/100ml较低浓度配体的(15.40±1.03)金氏单位/100ml有进一步下降(P=0.008),Runx2和Osterix表达的变化趋势与ALP活性一致。茜素红染色显示高浓度配体作用后钙结节明显减少。结论明确了EphB6对BMSC成骨分化有负性调控作用。[关键词]ephrinreceptor;骨髓间充质干细胞;成骨分化;组织工程[中图法分类号][文献标志码]AEffectofEphB6onosteogen
4、esisofbonemarrowstemcells:apreliminarystudyLiSongtao,LiuYong,SongLei,DongShiwu,DiNing,XuYuan,LiDianwei,ZhouQiang(DepartmentofOrthopaedics,SouthwestHospital,ThirdMilitaryMedicalUniversity,Chongqing400038,China)[Abstract]ObjectiveTodetecttheexpressiono
5、fEphB6duringosteogensisofbonemarrowstemcells(BMSC).AndapreliminarystudyofligandstimulatedEphB6onosteogensiewasperformed.MethodsTissueengineeredbonesestablishedviabiphasicseedingweredividedintoosteogenesis5-day(groupA),osteogenesis14-day(groupB),non-i
6、nduced5-day(groupC),non-induced14-day(groupD).Activityofalkalinephosphatase(ALP),mRNAofRunx2,OsterixandEphB6werequantitated.ThensolubleephrinB1wasadministratedforBMSCandRunx2wasquantitated.ThensolubleephrinB1-Fcwasadministratedinallgroupswiththesamet
7、estitems.ResultsALP(7.32±2.02King’sUnit/100mL),Runx2(3.4426±0.2585)andOsterix(2.4823±0.3292)reachedthepeakingroupB,andincreasedingroupA.EphB6wasdown-regulatedingroupAandB,andwaslowestinB(0.7543±0.0235).AftertheadministrationofsolubleephrinB1-Fc,EphB6
8、wasup-regulated(P<0.05).ALPactivitydecreased(P<0.05),andwaslessinhigh-doseephrinB1(12.20±1.30King’sUnit/100mL)comparedwithlow-doseephrinB1(15.40±1.03King’sUnit/100mL),P=0.008.similartrendswereobservedinRunx2andOsterix.Alizarinredstainingsuggestedanev