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1、铁调素对RAW264.7向破骨细胞早期分化及晚期分化的影响#王啸，张鹏，汪升，沈光思，徐又佳**510152025303540（苏州大学附属第二医院骨科）摘要：目的探索梯度剂量铁调素干预小鼠单核细胞系RAW264.7后，向破骨细胞前期分化及后期分化能力的差异。方法铁调素以梯度浓度干预RAW264.7，24h后以CCK8法检测各组细胞增殖能力。各组细胞均匀接种至噬骨馅窝板中，在含核因子kB受体活化因子配体（RANKL）培养基的环境下，梯度浓度铁调素干预RAW264.7，早期分化组细胞干预3d后行抗酒石酸酸性磷酸
2、酶（TRAP）染色；晚期分化组RANKL诱导3d，再以梯度浓度铁调素干预3d，6d后行TRAP染色。各组在干预终点时以Trizol消化细胞并冲洗晾干噬骨馅窝板，镜下观察噬骨馅窝吸收情况；消化下的细胞行q-PCR检测破骨相关基因表达差异。结果铁调素在0.4~1.6μM范围内能促进破骨细胞分化，早期分化各浓度组TRAP阳性细胞数及噬骨馅窝数无统计学差异，而晚期分化各组细胞分化有统计学差异。q-PCR结果与TRAP染色结果一致。结论铁调素对RAW264.7向破骨细胞晚期分化有促进作用，且呈剂量依赖性，而对早期分化影响甚
3、微。关键词：实验外科学；铁调素；破骨细胞；骨质疏松中图分类号：R-681TheinfluenceofhepcidinonearlyandterminaldifferentiationofRAW264.7monocytesintoosteoclastsWANGXiao,ZHANGPeng,WANGSheng,SHENGuangsi,XUYoujia(DepartmentofOrthopaedics,TheSecondAffiliatedHospitalofSoochowUniversity,SuZhou215004
4、)Abstract:ObjectiveToexploretheinfluenceofgradeddosesofhepcidinondifferentiationofmouseRAW264.7cellsintoosteoclasts,especiallyontheearlyorternimaldifferentiation.MethodsRAW264.7cellswereinterveredwithgradeddosesofhepcidininthepresenceofreceptoractivatorofNF-k
5、Bligand(RANKL).CellviabilitywastestedwithCCK8after24h,andthenreplatedintoosteoassayplates.Tostudytheeffectontheearlydifferentiation,RAW264.7cellsweretreatedwithindicatedconcentrationsofhepcidinfor3d.Ontheternimaldifferentiation,cellswereculturedinthepresenceo
6、fRANKLfor3dandthentreatedwithhepcidinforanother3d.,thenstainedwithtartrate-resistantacidphosphatase(TRAP).Numbersofpitsweremeasured.Geneexpressionswereexaminedwithq-PCR.ResultsHepcidinat0.4~1.6μMmightstimulateosteoclaststerminaldifferentiation,nottheearlydiff
7、erentiation,whichwasindicatedwiththeresultsofTRAPstainingandresorptionpitassay.ThegeneexpressionsofTRAP,CTKandCTRwereincorrespondencewiththeconsequencesofTRAPstainingandresorptionpitassay.ConclutionAsmalldoserangeofhepcidincouldsignificantlyfacilitatetheterni
8、maldifferentiation,nottheearlydifferentiation,ofRAW264.7cellsintoosteoclasts.Thiseffectwasdosedependent.Keywords:Experimentalsurgery1;Hepcidin2;Osteoclasts3;Osteoporosis0引言骨质疏松是以低骨量，骨组织微观
