糖尿病与氧化应激在颈动脉粥样硬化大鼠内皮功能障碍中的作用研究.docx

.论著糖尿病与氧化应激在颈动脉粥样硬化大鼠内皮功能障碍中的作用研究盛冲霄，黎红华，崔敏，汪志忠，李敏【摘要】目的探讨糖尿病与氧化应激在颈动脉粥样硬化(CAS)大鼠内皮功能障碍中的作用。方法2014年610月选取SPF级健康雄性WiSlar大鼠30只，采用随机数字表法分为对照组、CAS组、糖尿病CAS组，每组10只。CAS组、糖尿病CAS组大鼠给予高脂饲料喂养结合维生素1)3注射液60万U/kg一次性腹腔注射建立CAS模型。糖尿病CAS组大鼠第5周时给予链脱佐菌素(STZ)45mg/kg腹腔注射建立糖尿病模型。对照组大鼠给予基础饲料喂养结合09%氯化钠溶液腹腔注射。15周后处死大鼠，分离颈动脉，截取约3mm新鲜颈动脉环，HV-4离体组织器官恒温灌流系统中加入不同浓度(10-、10-°、10-7、10-6、io-'o-4mo"L)乙酰胆碱，记录颈动脉环张力(最大舒张度)。苏木素-伊红(HE)染色观察颈动脉组织，免疫组织化学SP法染色并检测颈动脉亲环素A(CyPA)、Kruppel样转录因子2(KLF2)、内皮型一氧化氮合醒(eNOS)平均光密度。结果HV-4离体组织器官恒温灌流系统中加入不同浓度乙酰胆碱后各组大鼠颈动脉环均出现舒张反应。CAS组、糖尿病CAS组大鼠10-、10-。、10-7、IO"、io、IOTmol/L乙酰胆碱颈动脉环张力较对照组降低(P<0.05)：糖尿病CAS组大鼠10-°、10-7、106、i()_5、IOTmOl/L乙酰胆碱颈动脉环张力较CAS组降低(P<0.05).HE染色示对照组大鼠颈动脉内膜完整光滑，内皮细胞以及平滑肌细胞排列整齐，无增生；弹性纤维连续无中断。CAS组、糖尿病CAS组大鼠颈动脉可见内膜断裂，中层钙化斑块形成，弹性层失去连续性。CAS组、糖尿病CAS组大鼠颈动脉CyPA平均光密度较对照组升高，KLF2、eNOS平均光密度较对照组降低(P<0.05);糖尿病CAS组大鼠颈动脉CyPA平均光密度较CAS组升高，KLF2平均光密度较CAS组降低(P<0.05).结论糖尿病与氧化应激参与了CAS大鼠内皮功能功能障碍的形成，其机制可能与CyPA表达水平增加有关。【关键闻】颈动脉疾病：糖尿病：氧化应激：内皮功能障碍；亲环素A【中图分类号】R743.1【文献标识码】doi;10.3969j.issn.1007-9572.2016.09.006盛冲霄，黎红华，崔敏，等.糖尿病与氧化应激在颈动脉粥样硬化大鼠内皮功能障碍中的作用研究J.中国全科医学,2016,19(9):1015-1020.ShengCX,LiHH.CuiM,etal.ImpactofdiabetesandoxidativestressonendothelialdysfunctionofcarotidatherosclerosisratsJ.ChineseGeneralPractice.2OI6,19(9):1015-1020.基金项目：湖北省自然科学基金资助项目(2013CFB428)作者单位：430070湖北省武汉市，广州军区武汉总医院神经内科通信作者：黎红华，430070湖北省武汉市，广州军区武汉总医院神经内科：E-mail:LihOnghUa567aliyun.COm9TakaseB,MasakiN,HattoriH,etal.UsefulnessofautomaticQTdispersionmeasurementtordetectingexercise-inducedmyocardialischemiaJ.AnadoluKardiyolDerg,2009,9(3):189-195.IOBonominiMP,AriniPD.GonzalezGE,etal.ThealIometricmodelinchronicmyocardialinfarctionJ.TheorBiolMedModel.2012,11(9):15.IIEslamiV.SafiM,TaherkhaniM,etal.EvaluationofQT.QTdispersion,andT-wavepeaktoendtimechangesafterprimarypercutaneouscoronaryinterventioninpatientspresentingwithacuteST-elevationmyocardialinfarctionJ.JInvasiveCardiol,2013,25(5):232-234.12贺红，王利青，王彩霞，等.ST段抬高型心肌梗死患者择期PCI术后左心功能及血尿酸水平的研究J.疑难病杂志，2011,10(8):572.13KorantzopoulosP,LelsasKP,ChristogiannisZ,elal.Exercise-inducedrepolarizationchangesinpatientswithstableCorOnalyarterydiseaseJ.AmJCardiol,2011,107(1):37-40.14HaarinarkC,GraffC,AndersenMP.etal.ReferencevaluesofelectrocardiogramrepolarizationvariablesinahealthypopulationJ.JElectrocardiol,2010,43(1):31-39.15任春霖，许浩.平板运动试验DUke评分对老年冠心病诊断价值J.中国煤炭工业医学杂志，2014,17：192-195.16.QaseemA1FihnSD,WilliamsS.etal.Diagnosisofstableischemicheartdisease:summaryofaclinicalpracticeguidelinefromtheAmericanCollegeOfPhysiciansZAmericanCollegeofCardiologyFoundationZAmericanHeartAssociationZAmericanAssociationforThoracicSurgeryZPreventiveCardiovascularNursesAssociation/SocietyofThoracicSurgeonsJ.AnnIntermMed,2012,157(10):729-734.(收稿日期：2015-06-16;修回日期：2016-01-06)(本文编辑：陈素芳)ImpactofDiabetesandOxidativeStressonEndothelialDysfunctionofCarotidAtherosclerosisRatsSHENGChong-xiao,LIHong-hua,CUIMin,etal.DepartmentofNeurology,WuhanGeneralHospitalofGuangzhouMiIiIaryAreaCommand,Wuhan430070,China(AbstractObjectiveToobservetheimpactofdiabetesandoxidativestressonendothelialdysfunctionofcarotidatherosclerosis(CAS)rats.MethodsFromJunetoOctoberin2014,30healthymaleSPF-gradeWistarratsweredividedintocontrolgroup,CASgroupanddiabeticCASgroupwith10ratsineachgroupbyrandomtablemethod.CASgroupanddiabeticCASgroupweregivenintraperitonealinjectionofasingledoseOfVitaminD,by600.000Uperkgbodyweightwithhigh-fatdiet.Inthefifthweek,diabeticCASgroupwasgivenintraperitonealinjectionofadoseofStreptozocinby45nigperkgbodyweight.Controlgroupwasfedwithbasicdiet.After15weeks,theratsweresacrificed,and3mmfreshcarotidarterialringwassectionedfromeachoftheseparatedcarotidarteries.Thecarotidarteryringtensionwasmeasuredbyaddingdifferentconcentrationsofacetylcholine(10-',10-,10-7,10-6,10-5and10molL)intoHV-4outbodytissueorganconstanttemperatureperfusionsystem.Carotidarterytissuewasobsen,edbyHematoxylinandEosinstaining,andtheaverageopticaldensityofCyPA,KLF2andeNOSwasmeasuredbyimmunohistochemistrySPmethod.ResultsAfter(headditionofdifferentconcentrationsofacetylcholineintoHV-4outbodytissueorganconstanttemperatureperfusionsystem,thecarotidarterialringsofallthethreegroupshadrelaxantresponses.CASgroupanddiabeticCASgroupwerelowerthancontrolgroupincarotidarterialringtensionwhenacetylcholinewasgivenby10-oJ08,10-7,10-6,10-5and10molL(P<0.05).DiabeticCASgroupwaslowerthanCASgroupincarotidarterialringtensionwhenacetylcholinewasgivenbyIO-8,10-',IO-6,10-5and10-4molL(P<0.05).HEstainingshowedthatcarotidarteryintimaofcontrolgroupwascompleteandsmoothwithendothelialcellsandsmoothmusclecellsinalignment,elasticfiberscontinuouswithoutbreakoff,andnohyperplasia.InCASgroupanddiabeticCASgroup,intimabreakage,fbnationofmedialcalcificationplaquesandlossofelasticlayercontinuitywereobserved.CASgroupanddiabeticCASgroupwerehigherintheaverageopticaldensityofCyPAandlowerintheaverageopticaldensityofKLF2andeNOSthancontrolgroup(P<0.05).DiabeticCASgroupwashigherintheaverageopticalde