Wntβ-catenin信号通路在氟致大鼠成骨细胞分化中的作用.docx

Wntβ-catenin信号通路在氟致大鼠成骨细胞分化中的作用
摘要：
氟化物是一种常见的工业物质，在人们的日常生活中也难以避免与氟化物接触。然而，过量摄入氟化物会对骨骼健康造成不良影响。本文采用大鼠成骨细胞培养模型，通过检测Wnt/β-catenin信号通路的变化，研究氟致大鼠成骨细胞分化的机制。结果显示，氟化物显著抑制了Wnt/β-catenin信号通路的活性，进而影响了大鼠成骨细胞的分化和成骨能力。本研究提供了在氟化物中毒情况下探究成骨细胞分化机制的新思路。
关键词：氟化物；Wnt/β-catenin信号通路；成骨细胞分化
Introduction
Fluorideiswidelydistributedintheenvironmentandiscommonlyusedinindustrialproducts,toothpaste,andwatersources.Overexposuretofluoridecancauseavarietyofhealthproblems,includingdentalandskeletalfluorosis.Skeletalfluorosisisabonediseasethatiscausedbyexcessivefluorideintakeandcanleadtoreducedbonedensity,brittlebones,anddeformities.Themechanismbywhichfluorideaffectsbonemetabolismisstillunclear,butrecentstudieshavesuggestedthattheWnt/β-cateninsignalingpathwayisinvolved.
TheWnt/β-cateninsignalingpathwayisahighlyconservedpathwaythatregulatesmanyaspectsofembryonicdevelopment,tissuehomeostasis,andstemcellrenewal.Thispathwayiscriticalforbonedevelopmentandhomeostasis,anditsactivationcanpromoteosteoblastdifferentiationandboneformation.β-cateninisakeycomponentofthispathwayandisresponsiblefortransducingtheWntsignaltothenucleus,resultingintheactivationofWnttargetgenes.Inthisstudy,weinvestigatedtheroleoftheWnt/β-cateninsignalingpathwayinfluoride-inducedosteoblastdifferentiation.
MaterialsandMethods
PrimaryosteoblastswereisolatedfromthecalvariaofnewbornSprague-Dawleyratsandculturedinα-MEMmediumsupplementedwith10%FBSandantibiotics.Thecellsweretreatedwithsodiumfluoride(NaF)atconcentrationsof0,1,5,and10mMfordifferenttimeperiods.TheexpressionlevelsofkeygenesinvolvedintheWnt/β-cateninpathwayweremeasuredbyqPCR,westernblotting,andimmunofluorescencestaining.Alkalinephosphatase(ALP)activityandmineralizationassayswereperformedtoevaluatetheosteogenicdifferentiationofthecells.
Results
TreatmentwithNaFsignificantlyinhibitedtheosteogenicdifferentiationofprimaryosteoblasts,asevidencedbydecreasesinALPactivityandmineralization.qPCRanalysisrevealedthattheexpressionlevelsofkeygenesinvolvedintheWnt/β-cateninpathway,includingβ-cate