Sclerostin Stimulates Osteocyte Support of Osteoclast Activity by a RANKL-Dependent Pathway 英文参考文献.docVIP

SclerostinStimulatesOsteocyteSupportofOsteoclast ActivitybyaRANKL-DependentPathway AsiriR.Wijenayaka1,MasakazuKogawa1,HuiPengLim1,LyndaF.Bonewald2,DavidM.Findlay1, GeraldJ.Atkins1* 1BoneCellBiologyGroup,DisciplineofOrthopaedicsandTrauma,UniversityofAdelaide,andtheHansonInstitute,Adelaide,Australia,2UniversityofMissouri-Kansas CitySchoolofDentistry,DepartmentofOralBiology,KansasCity,Missouri,UnitedStatesofAmerica Abstract Sclerostinisaproductofmatureosteocytesembeddedinmineralisedboneandisanegativeregulatorofbonemassand osteoblastdifferentiation.Whileevidencesuggeststhatsclerostinhasananti-anabolicrole,thepossibilityalsoexiststhat sclerostin has catabolic activity. To test this we treated human primary pre-osteocyte cultures, cells we have found are exquisitely sensitive to sclerostin, or mouse osteocyte-like MLO-Y4 cells, with recombinant human sclerostin (rhSCL) and measuredeffectsonpro-catabolicgeneexpression.Sclerostindose-dependentlyup-regulatedtheexpressionofreceptor activatorofnuclearfactorkappaB(RANKL)mRNAanddown-regulatedthatofosteoprotegerin(OPG)mRNA,causingan increase in the RANKL:OPG mRNA ratio. To examine the effects of rhSCL on resulting osteoclastic activity, MLO-Y4 cells plated onto a bone-like substrate were primed with rhSCL for 3 days and then either mouse splenocytes or human peripheral blood mononuclear cells (PBMC) were added. This resulted in cultures with elevated osteoclastic resorption (approximately 7-fold) compared to untreated co-cultures. The increased resorption was abolished by co-addition of recombinantOPG.Inco-culturesofMLO-Y4cellswithPBMC,SCLalsoincreasedthenumberandsizeoftheTRAP-positive multinucleatedcellsformed.Importantly,rhSCLhadnoeffectonTRAP-positivecellformationfrommonoculturesofeither splenocytesorPBMC.Further,rhSCLdidnotinduceapoptosisofMLO-Y4cells,asdeterminedbycaspaseactivityassays, demonstrating that the osteoclastic response was not driven by dying osteocytes. Together, these results suggest that sclerostinmayhaveaca