Perturbation of adhesion molecule-mediated chondrocyte-matrix interactions by 4-hydroxynonenal binding implication in osteoarthritis pathogenesis 英文参考文献.docVIP

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Perturbation of adhesion molecule-mediated chondrocyte-matrix interactions by 4-hydroxynonenal binding implication in osteoarthritis pathogenesis 英文参考文献.doc

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Perturbation of adhesion molecule-mediated chondrocyte-matrix interactions by 4-hydroxynonenal binding implication in osteoarthritis pathogenesis 英文参考文献

El-Bikaietal.ArthritisResearchTherapy2010,12:R201 /content/12/5/R201 RESEARCH ARTICLE OpenAccess Perturbationofadhesionmolecule-mediated chondrocyte-matrixinteractionsby 4-hydroxynonenalbinding:implication inosteoarthritispathogenesis RanaEl-Bikai,MélanieWelman,YoranMargaron,Jean-Fran?oisC?té,LukeMacqueen,MichaelDBuschmann, HassanFahmi,QinShi,KarimMaghni,JulioCFernandes,MohamedBenderdour* Abstract Introduction:Objectivesweretoinvestigatewhetherinteractionsbetweenhumanosteoarthriticchondrocytesand 4-hydroxynonenal(HNE)-modifiedtypeIIcollagen(ColII)affectcellphenotypeandfunctionsandtodeterminethe protectiveroleofcarnosine(CAR)treatmentinpreventingtheseeffects. Methods:HumanColIIwastreatedwithHNEatdifferentmolarratios(MR)(1:20to1:200;ColII:HNE).Articular chondrocyteswereseededinHNE/ColIIadduct-coatedplatesandincubatedfor48hours.Cellmorphologywas studiedbyphase-contrastandconfocalmicroscopy.Adhesionmoleculessuchasintercellularadhesionmolecule-1 (ICAM-1)anda1b1integrinatproteinandmRNAlevelswerequantifiedbyWesternblotting,flowcytometryand real-timereversetranscription-polymerasechainreaction.Celldeath,caspasesactivity,prostaglandinE2(PGE2), metalloproteinase-13(MMP-13),mitogen-activatedproteinkinases(MAPKs)andnuclearfactor-kappaB(NF-B) wereassessedbycommercialkits.ColII,cyclooxygenase-2(COX-2),MAPK,NF-B-p65levelswereanalyzedby Westernblotting.Theformationofa1b1integrin-focaladhesionkinase(FAK)complexwasrevealedby immunoprecipitation. Results:ColIImodificationbyHNEatMRapproximately1:20,stronglyinducedICAM-1,a1b1integrinandMMP-13 expressionaswellasextracellularsignal-regulatedkinases1and2(ERK1/2)andNF-B-p65phosphorylationwithout impactingcelladhesionandviabilityorColIIexpression.However,ColIImodificationwithHNEatMR approximately1:200,alteredchondrocyteadhesionbyevokingcelldeathandcaspase-3activity.Itinhibiteda1b1 integrinandColIIexpressionaswellasERK1/2 andNF-B-p65phosphorylation,but,incontrast,markedlyelicited PGE release,COX-2expressionandp38MAPKphosphorylation