重组人白介素-10抑制晚期糖基化终产物诱导的大鼠血管平滑肌细胞和血管

Received2002208212Accepted2002211227ThisprojectwassupportedbygrantsfromtheFoundationofGuangzhouScienceandTechnologyCommittee,P.R.China(JB00000448165).3Correspondingauthor.Tel:[phone];Fax:[phone];E2mail:ls36@zsu1edu1cnResearchPaperRecombinanthumaninterleukin210inhibitsproliferationofvascularsmoothmusclecellsstimulatedbyadvancedglycationendproductsandneointimahyperplasiaaftercarotidinjuryintheratOUYANGPing,PENGLi2Sheng,YANGHong,PENGWen2Lie,WUWen2Yan,XUAn2Long3DepartmentofBiochemistry,SchoolofLifeScience,SunYat2senUniversity,Guangzhou510275,ChinaAbstract:Thepurposesofthisstudywastodeterminetheeffectsofrecombinanthumaninterleukin210(rhIL210)onpro2liferationofvascularsmoothmusclecells(VSMCs)stimulatedbyadvancedglycationendproducts(AGE)andneointimahy2perplasiaafterratcarotidarterialinjury.RataorticVSMCswereculturedandtreatedwithrhIL210orAGErespectively,andthenco2treatedwithrhIL210andAGE.ProliferationofVSMCswasquantifiedbycolormetricassay.Cellcycleanalysiswasperformedbyflowcytomertry.Sprague2DawleyratsweretreatedwithrecombinanthumanIL210(rhIL210)for3daftercarotidarteriesinjury.Theratioofneointimatomediaareaatthesiteofarterialinjurywasmeasured28dafterballoonin2jury.Thep44/42MAPKactivitywasevaluatedbytheimmunoblottingtechniqueusinganti2p44/42phospho2MAPKanti2body.Comparedtocontrol,AGEstimulatedVSMCsproliferation.rhIL210alonehadnoeffectonVSMCsgrowth.WithAGEstimulation,rhIL210,atdoseaslowas10ng/ml,inhibitedVSMCsgrowth(P0105).ThecellnumberinG0/G1phaseofAGEandrhIL210co2treatmentgroupwashigherthanthatofAGEtreatmentalone(P0101)byflowcytometryanalysis.Comparedwiththecontrolgroupofneointimahyperplasiainrats,theratioofneointimatomediaareaofrecombi2nanthumanIL210groupwasreducedby45%(P0101).Thep44/42MAPKactivitywassignificantlyenhancedbyAGE.TheAGEeffectswereopposedbyrhIL210.Theanti2inflammatorycytokinerhIL210inhibitsAGE2inducedVSMCsproliferation.RecombinanthumanIL210alsoinhibitedneointimahyperplasiaaftercarotidarteryinjuryinrats.TheresultssuggestthepossibilitythatrecombinanthumanIL210,asapotentialtherapeuticapproach,preventsneointimalhyperplasia.Keywords:pathology;interleukin210;muscle;smooth;vascular;advancedglycationendproducts;neointimahyperplasia210,,,,,3,510275:10(rhIL210)(AGE)SD,MTS/PES;;p44/42MAPKp44/42MAPK,rhIL210:(1)AGE,AGE(P0105)rhIL210(P0105)AGE,100ng/mlrhIL210(P0105)(2),rhIL210AGEVSMCG0/G1,(P0101)(3)AGEp44/p42MAPK,rhIL210(P01001)(4),rhIL210/45%(P0101)rhIL210AGE:;10;;;;;:Q463821ActaPhysiologicaSinica,April25,2003,55(2):128-134©1995-2005TsinghuaTongfangOpticalDiscCo.,Ltd.Allrightsreserved.Accumulatingevidencesuggeststhatatherogenesisrepresentsanexaggeratedinflammatoryresponsetovascularinjury.Aftervesselinjury,monocytes,platelets,andlymphocytesadheretothevesselwallandreleaseanarrayofcytokinesandpeptidegrowthfactors.Thesegrowth2regulatorysubstancesbindtotheirrespectivereceptorsandtransducesignalsthatmayinfluencethephenotypeandgrowthofvascularsmoothmusclecells(VSMCs),thuspromotingdevel2opmentofadvancedfibroproliferativelesions[1].Smoothmusclecellsinatheromatouslesions,butnotinnormalhumanarteries,expressthehumanleuko2cyteantigenDR(HLA2DR)[2].Encodedbythemajorhistocompatibilitycomplex(MHC),theHLA2DRglycoproteinparticipatesinantigenpresentationtoTcells.Assuch,alocalimmuneresponse,assuggestedbyTlymphcyte2VSMCsinteractions,maycontributetoatherogenesis.Manipulationoftheanti2inflamma2toryproductsofTlymphocytesmayprovideathera2peuticstrategyagainstpathologicvascularremodeling.Intimalhyperplasiaisauniversalresponseofthear2terialwalltoballooninjuryandthemainmechanismofrestenosis[3]afterstentimplantation[4],atechniqueusedforthemajorityofpercutaneouscoronaryinter2ventions.Inflammatorycellsplayakeyroleinpostin2juryintimalhyperplasia.Inthedaysafterballoonan2gioplasty[5]orstenting[6],monocytesarerecruitedattheinjurysite,wheretheybecomeactivatedmacrophages.Themagnitudeofmacrophageinfiltra2tioninstentedlesionsiscorrelatedtosubsequentinti2malgrowth[7].Modificationofproteinsbylong2termincubationwithglucoseleads,throughtheformationofearlystageproductssuchasSchiffbaseandAmodorirear2rangementproducts,totheformationofadvancedgly2cationendproducts(AGE).AGEcanalterproteinfunctionanddisturbcellularmetabolism.Theoccur2renceofadvancedglycationendproductsisconsideredtodayasthemainpathogenicagentofatherosclero2sis[8].Interleukin210(IL210)isananti2inflammatorycy2tokinewithapowerfulinhibitoryeffectonmonocytes.TheprincipalroutinefunctionofIL210appearstolim2itandultimatelyterminateinflammatoryresponses[9].WestudiedtheeffectofrecombinanthumanIL210(rhIL210)onproliferationofratVSMCsbyAGE,andneointimahyperplasiaaftercarotidarteriesinjuryinrat.1MATERIALSANDMETHODS111Materials.Hanksbalancedsaltsolution,DulbeccosmodifiedEaglesmedium(DMEM),phosphate2bufferedsaline(PBS),fetalbovineserum(FBS),L2glutamineandantibiotic(penicillinG10000U/ml,streptomycinsulfate10000mg/ml)wereobtainedfromGibcoBrl(GrandIsland,NY,USA).Propidiumiodide(PI),col2lagenaseandelastasewereobtainedfromSigmaChemical(StLouis,Mo,USA).RecombinanthumanIL210pro2teinwasobtained