gdf-5在小鼠骨髓基质干细胞诱导软骨分化过程中的实验研究

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1、华中科技大学博士学位论文够抑制GDF-5诱导MSC向软骨方向的分化。Alcian染色结果显示对照组阴性，GDF-5组呈广泛的蓝染，GDF-5+1-庚醇组染色较GDF-5组明显减弱，但仍成蓝色，说明1-庚醇能够抑制蛋白多糖的分泌。Westernblotting结果表明Cx43蛋白相对量分别为：对照组0.326±0.004；GDF-5组0.582±0.006；GDF-5+1-庚醇组0.576±0.014，GDF-5和GDF-5+1-庚醇组与对照组相比有显著性差异，而GDF-5与GDF-5+1-庚醇组差异无显著性意义，说明GDF-5能够上调Cx43蛋白的表达，而1-庚醇Cx43的不产生作用

2、。结论GDF-5能够定向诱导小鼠骨髓基质干细胞向软骨方向分化，软骨发生早期诱导Cx43蛋白高表达，继而缝隙连接细胞间通讯（GJIC）功能增强，从而促发其软骨分化，Cx43介导的GJIC在GDF-5诱导软骨分化中发挥着很重要的作用。【关键词】GDF-5；软骨；缝隙连接；阻断剂；细胞间通讯；小鼠；骨髓基质干细胞VI华中科技大学博士学位论文ResearchinMechanismsofGrowthandDifferentiationFactor5PromotingChondrogenicDifferentiationofMouseBoneMarrowMesenchymalStemCellsD

3、epartmentofOrthopedics,UnionHospital,TongjiMedicalCollegeofHuazhongUniversityofScienceandTechnologyPh.D.Candidate:ZhangYukunSupervisor:YangShuhuaABSTRACTPartⅠExpressionandSignificanceofGDF-5DuringLimbSkeletalDevelopmentofMiceObjectiveToinvestigatetheexpressiontrendandeffectofgrowthdifferentiati

4、onfactor5(GDF-5)duringlimbskeletaldevelopmentofmice.MethodsMouseembryoswereobtainedfrommiceofKunmingstrainsprovidedbyDepartmentofExperimentalAnimalofTongjiMedicalCollegeofHuazhongScienceandTechnologyUniversity.Thedayavaginalplugwasfoundwasconsideredasembryonicday0.5(E0.5).DuringE11.5-15.5embryo

5、swereremovedfromthepregnantfemalemiceandlimbbudsweretakenoutofthemtobekeptinreserveforusewithophthalmicscissors.TheexpressionofGDF-5mRNAandproteinofmousefetallimbbudsweredetectedinembryonicdays11.5-15.5(E11.5-15.5)byRT-PCRandWesternblottingrespectively.Results①ExpressionofGDF-5mRNAinmouseembryo

6、niclimbbuds:A219bpofspecificelectrophoreticstripwasobservedineverylimbbudandaveragelightdensityratioofGDF5/β-actinwas0.521±0.023,1.210±0.014,1.221±0.008,0.556±0.014,0.510±0.019VII华中科技大学博士学位论文respectivelyfromE11.5toE15.5.TheresultsdemonstratethattheexpressionofGDF-5mRNAwasconstantduringearlystag

7、eofdevelopmentalskeletogenesisandbeganwithembryosE11.5,highlightedatembryosE12.5andE13.5,subsequentlydroppedatembryosE14.5andE15.5.Therewasverysignificantdifference(p<0.01)inaveragelightdensityratioofGDF-5/β-actinbetweenE12.5-13.5