smad7在sox9增强bmp2成软骨效应中的作用

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1、Smad7在Sox9增强BMP2成软骨效应中的作用赵辰，黄伟，梁熙，廖军义，周年，胡宁，赵智，简长春（400016重庆，重庆医科大学附属第一医院骨科）[摘要]目的探讨Smad7在Sox9增强BMP2成软骨效应中的作用和机制。方法小鼠骨髓间充质干细胞（C3H10T1/2）为种子细胞，重组腺病毒AdBMP2和AdSox9感染细胞，AdGFP感染细胞为对照。运用Real-TimePCR，免疫组化和Westernblot法分别检测感染后各组Smad7转录水平和蛋白质表达情况。Real-TimePCR检测与Smad7相关因子MMP13与OPN表达。结果AdBMP2和AdSox9联合感染

2、细胞7、11d时，Smad7mRNA和蛋白质表达水平均明显低于AdBMP2单独感染组(P<0.05)，免疫组化染色BMP2+Sox9组Smad7染色明显弱于BMP2单用组。同时BMP2+Sox9组中与软骨最终成熟因子OPN与MMP13的表达均低于BMP2单用组(P<0.05)。结论Sox9可抑制BMP2诱导下Smad7的高表达，解除其对BMP2成软骨的抑制作用，并抑制Smad7相关因子MMP13与OPN表达，阻止软骨细胞最终成熟骨化，保持软骨发育与正常状态。[关键词]软骨；Sox9；BMP2；Smad7[中图法分类号][文献标志码]ATheeffectofSmad7inSox

3、9PotentiatesBMP2-InducedChondrogenicDifferentiationZhaoChen1，HuangWei1，HuNing1，LiaoJunyi1，ZhouNian1，LiangXi1，ZhaoZhi1，JianChangchun1（DepartmentofOrthopedics，TheFirstAffiliatedHosphalofChongqingMedicalUniversity，Chongqing400016，China）[Abstract]Objective：WehavefoundSox9canenhancechondrogenicd

4、ifferentiationinducedbyBMP2,howeverthemechanismremainunknown.BytestingExpressionofSmad7anditsrelatedfactorsMMP13andOPN,wecanfindwhetherSox9cancooperatewithBMP2byinhibitingSmad7.Method：MouseembryonicbonemarrowMSCC3H10T1/2cellswereinfectedwithrecombinantadenovirusexpressingBMP2,Sox9for3-11day

5、s,withcellsinfectedwiththeadenoviruscarryingGFPgeneasthecontrol..ThemRNAexpressionoftheSmad7anditsrelatedfacotorsMMP13andOPNweredeterminedbyreal-timePCR.TheexpressionofSmad7proteinwasassayedbyimmunohistochemicalstainingandWesternblotanalysis.Result：BMP2increaseexpressionofSmad7in3d,7d,11d.O

6、verexpressionofSox9effectivelyreduceBMP2-inducedexpressionoftheSmad7inbothmRNAandproteinlevel.MMP13andOPNarealsorepressedbyoverexprssionofSox9.Conclusion：Co-expressionofBMP2andSox9canreduceexpressionofSmad7.OverexpressionofSox9duringBMP2inducedchondrogenicdiffrentationcouldnotonlyenhancethe

7、chondrogenicalfactorbutalsorepresstheMMP13andOPNleadtobyinhibitexpressionofSmad7.［Keywords］BMP2；Smad7；Sox9；CartilgeSupportedbytheGeneralProgramofNationalNaturalScienceFoundationofChina(31070875，8371972).Correspondingauthor:HuangWei,E-mail:huangwei68@263.