持续牵张促进小鼠骨髓间充质干细胞成骨向分化机制的研究

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1、口腔医学研究2015年8月第31卷第8期775持续牵张促进小鼠骨髓间充质干细胞成骨向分化机制的研究刘小雅肖文林于保军(青岛大学附属医院黄岛院区口腔颌面外科山东青岛266555)[摘要]目的：研究p38MAPK在持续性机械牵张力促进C57BL／6J小鼠骨髓间充质干细胞成骨分化过程中的作用机制。方法：C57BL／6J小鼠BMSCs被随机分为对照组和牵张力阻断组。牵张力阻断组预先用p38MAPK特异性阻断剂SB203580处理1h，后用Flexercell加力仪加载0．5Hz，0．8的牵张力。对照组不做特

2、殊处理。分别对两组细胞施加0、30rain和60min的牵张力。采用Westernblot检测P—p38MAPK蛋白的表达情况，Realtime—PCR检测成骨基因ALP、COLI、OCNmRNA的表达变化。结果：C57BL／6J小鼠BMSCs传代后细胞生长状态稳定，呈梭形，具有多向分化潜能。Realtime—PCR结果显示对照组BMSCs中ALP、COLI、OCNmRNA表达在不同时间点(30rain与0min，60rain与30min)间差异均有统计学意义(P

3、、OCN的表达量均明显高于同一时间点(30rain和60min)牵张力阻断组(P<0．05)。Westernblot结果显示，对照组内BMSCs中P—p38MAPK的蛋白水平在不同时间点间(30min与0min，60min与30min)差异均有统计学意义(P<0．05)；对照组P—p38MAPK蛋白的表达量均明显高于同一时间点(30min和60min)牵张力阻断组(P

4、键词]持续牵张力骨髓间充质干细胞成骨细胞分化p38MAPK信号通路[中图分类号]R329．2[文献标识码]A[文章编号]1671—7651(2015)O8一O775一O4MechanismofOsteogenesisofBMSCsunderContinuousMechanicalTensionForce．L儿JXiao—n，jAoWB—lin，yUBao-jun．DepartmentofStomotology，theAffiliatedHospitalofQingdaoUniversity．Qing

5、dao266555[Abstract]Objective：Tostudytheroleofp38MAPKsignalpathwayinregulatingosteogenicdifferentiationofC57BL／6JmouseBMSCsunderthecontinuousmechanica1tensionforce．Methods：TheBMSCsofprimarycultureweredividedrandomlyintocontrolgroupandtensioninhibitorgro

6、up．Thetensioninhibitorgroupwaspretreatedfor1hourwithSB203580whichwasoneofp38MAPKpathwayspecificinhibitor，andthenloadedwithmechanicalstimulationbyFlexercel1．Thetensionforce(O．5Hz，0．8)wasinflictedtObothgroupsatO，30and60rain，re—spectively．p38MAPKandP—p38M

7、APKproteinweredetectedbyWesternblot。andtheexpressionofosteogenicgenesALP，COLI，0CNweredetectedbyrealtime—PCR．Results：C57BL／6JmouseBMSCsgrewwellwithtypi—calshuttleshapeandcapacitiesofmulti—differentiati0n．Rea1time—PCRandWesternblotresultsshowedthattheexp

8、ressionofALP，COLI，OCNmRNAandP—p38MAPKproteininthecontrolgroupwassignificantdifferenta—mongdifferentloadingtimepoints．ThemRNAandP—p38MAPKproteinexpressionincontrolgroupwasmarked—lyhigherthanthoseinthetensioninhibitorgroupatdifferenttimep