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1、第四军医大学硕士学位论文目的:探讨RNA连接蛋白QKI对大鼠心肌成纤维细胞增殖及胶原合成的影响,进一步完善心肌纤维化的相关机制,为改善心机重构提供新的思路。方法:采用胶原酶消化,差速贴壁法原代培养SD乳鼠心肌成纤维细胞,建立AngII诱导CF增殖及胶原合成模型,运用western-blot法,RT-PCR法,流式细胞术观察过表达及干涉QKI对其的影响和p27在其中的表达变化。结果:在CF中血管紧张素II诱导了QKI表达时间依耐性的降低,在24h后降至谷底。过表达或干涉QKI后通过western-blot法及RT-PCR法检测了CF中PC
2、NA、COX-2及collagen1a/3a表达水平的变化,流式细胞术观察细胞周期,数据表明QKI-6的过表达能明显抑制CF的增殖,干涉QKI后观察到CF增殖信号表达增加,但可能降低了CF对外界刺激耐受性。QKI的变化对CF胶原合成的影响并不明显。通过PT-PCR法定量检测p27在CF增殖模型、QKI过表达及干涉情况下的RNA表达水平,发现在增殖模型及QKI过表达的情况下,p27与QKI的表达变化具有正相关性,而在干涉QKI的情况下,二者相关性差异不显著。结论:QKI通过MAPK-ERK信号通路作用于p27,对CF增殖发挥负性调节作用,
3、并可能影响到CF细胞对外界刺激的耐受性,但对胶原合成并无明显影响。关键词:QKI;血管紧张素II;心肌成纤维细胞;增殖;胶原合成-4-第四军医大学硕士学位论文QKIaffectsontheproliferationandcollagensynthesisofratcardiacfibroblastsCandidateformaster:LiRuo-yuSupervisor:Prof.ZhengQiang-sunDepartmentofCardiovascularMedicine,Tang-duhospital,FourthMilitar
4、yMedicalUniversity,Xi’an710032,ChinaAbstractToday'sworld,cardiovasculardiseaseseriouslyendangeringthehealthandsafetyofhumanlife,heartfailureisacommonexpressiontoendstageofcardiovasculardiseases.IntheUnitedStates,about400,000casesofheartfailurepatientswithnewlyissued,atot
5、alofabout3millionpeoplesufferingfromheartfailure,about20milliondeathseachyear.Greaterthan65-year-oldcrowd,thecauseofheartfailurehospitalizationamongthefirst.Theannualcostoftreatingheartfailureinmorethan10billionU.S.dollars.Heartfailureisconsideredthekeyissuesinthenewcent
6、ury,knownasthe"newepidemicofcardiovasculardisease."Myocardialremodelingisanimportantpathophysiologicalbasisofheartfailure,includingmyocardialcellsremodelingandmyocardialremodeling,themostimportantpartofmyocardialremodelingismyocardialfibrosis.Therefore,thereversalofmyoca
7、rdialfibrosisandimprovethepreventionandtreatmentofmyocardialremodelinginheartfailureisimportant.QKIisagroupofRNAbindingproteinrecentlydiscovered,encodedbytheqkinggene,itsrolein-5-第四军医大学硕士学位论文regulatingmyelinformationhasbeenwellknown.QKIinheartcellsalsohaveawiderangeofexp
8、ression,recentstudieshavedeterminedthatithascloseto1430presumedtargetmRNA,andtheseincludethep27,acellul