Profiling Insulin Like Factor 3 (INSL3) Signaling in Human Osteoblasts 英文参考文献.docVIP

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Profiling Insulin Like Factor 3 (INSL3) Signaling in Human Osteoblasts 英文参考文献.doc

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Profiling Insulin Like Factor 3 (INSL3) Signaling in Human Osteoblasts 英文参考文献

ProfilingInsulinLikeFactor3(INSL3)SignalinginHuman Osteoblasts AlbertoFerlin1,LisaPerilli1,LisaGianesello1,GiuseppeTaglialavoro2,CarloForesta1* 1Section of Clinical Pathology, Department of Histology, Microbiology and Medical Biotechnologies, University of Padova, Padova, Italy, 2Orthopaedic and TraumatologicalClinic,UniversityofPadova,Padova,Italy Abstract Background:YoungmenwithmutationsinthegenefortheINSL3receptor(Relaxinfamilypeptide2,RXFP2)areatriskof reduced bone mass and osteoporosis. Consistent with the human phenotype, bone analyses of Rxfp22/2 mice showed decreasedbonevolume,alterationsofthetrabecularbone,reducedmineralizingsurface,boneformation,andosteoclast surface.TheaimofthisstudywastoelucidatetheINSL3/RXFP2signalingpathwaysandtargetsinhumanosteoblasts. Methodology/Principal Findings: Alkaline phosphatase (ALP) production, protein phosphorylation, intracellular calcium, geneexpression,andmineralizationstudieshavebeenperformed.INSL3inducedasignificantincreaseinALPproduction, andWesternblotandELISAanalysesofmultipleintracellularsignalingpathwaymoleculesandtheirphosphorylationstatus revealed that the MAPK was the major pathway influenced by INSL3, whereas it does not modify intracellular calcium concentration.QuantitativeRealTimePCRandWesternblottingshowedthatINSL3regulatestheexpressionofdifferent osteoblastmarkers.Alizarinred-SstainingconfirmedthatINSL3-stimulatedosteoblastsarefullydifferentiatedandableto mineralizetheextracellularmatrix. Conclusions/Significance: Together with previous findings, this study demonstrates that the INSL3/RXFP2 system is involvedinbonemetabolismbyactingontheMAPKcascadeandstimulatingtranscriptionofimportantgenesofosteoblast maturation/differentiationandosteoclastogenesis. Citation:FerlinA,PerilliL,GianeselloL,TaglialavoroG,ForestaC(2011)ProfilingInsulinLikeFactor3(INSL3)SignalinginHumanOsteoblasts.PLoSONE6(12): e29733.doi:10.1371/journal.pone.0029733 Editor:LinMei,MedicalCollegeofGeorgia,UnitedStatesofAmerica ReceivedMay30