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1、Osterix基因敲除及过表达对小鼠脊椎骨量的影响*陈思旭,宗兆文,贾敏,华祥,沈岳,赵玉峰,郭庆山,刘道城(400042重庆,第三军医大学大坪医院野战外科研究所创伤科,创伤、烧伤与复合伤国家重点实验室)[摘要]目的研究小鼠Osterix基因敲除及过表达后对其脊椎骨量的影响及作用机制。方法将12周龄的野生型小鼠(WT)、Osterix基因敲除小鼠(Osterix-KO)和Osterix过表达转基因小鼠(Osterix-CA)分为3组(每组8只,均为雄性),分别采用X线摄像和HE染色观察小鼠腰椎骨量的变化,TRAP染色检测破骨细胞水平,免疫组织化学检测NF-κB受体活化因子配体(RAN
2、KL)、骨涎蛋白(BSP)和骨桥蛋白(OPN)表达水平,并将所得数据进行统计学分析。结果Osterix-KO小鼠在X线和HE染色表现为腰椎骨密度和骨量增加,破骨细胞数量显著减少(P<0.05),椎体中RANKL、BSP和OPN的表达水平显著降低(P<0.05),而Osterix-CA小鼠的脊椎X线、HE染色表现,及破骨细胞数量与WT小鼠无明显差异。结论Osterix基因敲除小鼠脊柱骨吸收活性减弱伴成骨细胞分化能力降低,进而发生骨量明显增加。[关键词]Osterix;脊椎;骨量[中图法分类号][文献标志码]AEffectTheroleofOsterixosterixknock-outo
3、rover-expressionioncontrollingbonevolumeinvertebralbody.inmiceChenSixu,ZongZhaowen,JiaMin,HuaXiang,ShenYue,ZhaoYufeng,GuoQingshan,LiuDaocheng(StateKeyLaboratoryofTrauma,,BurnandCombinedInjury,,DepartmentofTrauma,,InstituteofSurgeryResearch,,DapingHospital,,ThirdMilitaryMedicalUniversity,,Chongq
4、ing,,400042,,China)[Abstract]ObjectiveToinvestigatethechangesofbonevolumeinvertebralbodyandtheunderlyingpossiblemechanisminOosterixknock-out(Osx-KO)andconstitutive-activation(Osx-CA)mice.MethodsAtotalof24Malemalemiceweredividedintothree3groups,thatis,includingwildtype(WT),Osx-KOandOsx-CAgroups(
5、n=8each).X–-rayradiologyandHEstainingwereusedtoevaluatethechangeofbonevolume.TRAPstainingwasusedtoassesstheactivityofosteoclasts.Immunohistochemistrycalassay(IHC)wasusedtoexaminetheexpressionlevelsofRANKL,BSPandOPN.TheIHCandTRAPstainingresultsweresemi-quantitiveanalyzedbymanualcountingmethod.Re
6、sultsX–-rayexaminationandHEstainingshowedthatthebonedensityandbonevolumeinthelumbarvertebralbodywasincreasedsignificantlyinOSX-KOmice,whileTRAPstainingshowedthatthenumberofosteoclastswasdecreased(P<0.01),andIHCrevealedthattheexpressionlevelsofRANKL,BSPandOPNweredown-regualted(P<0.01)inOSX-KOmic
7、e.However,Nnoobviouschangeswasfoundinofthebonedensity,bonevolumeandthenumberofosteoclastswerefoundinOsterix-KOmicecomparedwiththeWTmice.ConclusionsOsterixknock-outmayweakenboneresorptionactivityandosteoblastdifferentiationinthelum