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1、第四軍醫(yī)大學(xué)碩士學(xué)位論文目的：探討RNA連接蛋白QKI對(duì)大鼠心肌成纖維細(xì)胞增殖及膠原合成的影響，進(jìn)一步完善心肌纖維化的相關(guān)機(jī)制，為改善心機(jī)重構(gòu)提供新的思路。方法：采用膠原酶消化，差速貼壁法原代培養(yǎng)SD乳鼠心肌成纖維細(xì)胞，建立AngII誘導(dǎo)CF增殖及膠原合成模型，運(yùn)用western-blot法，RT-PCR法，流式細(xì)胞術(shù)觀察過(guò)表達(dá)及干涉QKI對(duì)其的影響和p27在其中的表達(dá)變化。結(jié)果：在CF中血管緊張素II誘導(dǎo)了QKI表達(dá)時(shí)間依耐性的降低，在24h后降至谷底。過(guò)表達(dá)或干涉QKI后通過(guò)western-blot法及RT-PCR法檢測(cè)了CF中PC
2、NA、COX-2及collagen1a/3a表達(dá)水平的變化，流式細(xì)胞術(shù)觀察細(xì)胞周期，數(shù)據(jù)表明QKI-6的過(guò)表達(dá)能明顯抑制CF的增殖，干涉QKI后觀察到CF增殖信號(hào)表達(dá)增加，但可能降低了CF對(duì)外界刺激耐受性。QKI的變化對(duì)CF膠原合成的影響并不明顯。通過(guò)PT-PCR法定量檢測(cè)p27在CF增殖模型、QKI過(guò)表達(dá)及干涉情況下的RNA表達(dá)水平，發(fā)現(xiàn)在增殖模型及QKI過(guò)表達(dá)的情況下，p27與QKI的表達(dá)變化具有正相關(guān)性，而在干涉QKI的情況下，二者相關(guān)性差異不顯著。結(jié)論：QKI通過(guò)MAPK-ERK信號(hào)通路作用于p27，對(duì)CF增殖發(fā)揮負(fù)性調(diào)節(jié)作用，
3、并可能影響到CF細(xì)胞對(duì)外界刺激的耐受性，但對(duì)膠原合成并無(wú)明顯影響。關(guān)鍵詞：QKI；血管緊張素II；心肌成纖維細(xì)胞；增殖；膠原合成-4-第四軍醫(yī)大學(xué)碩士學(xué)位論文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-第四軍醫(yī)大學(xué)碩士學(xué)位論文regulatingmyelinformationhasbeenwellknown.QKIinheartcellsalsohaveawiderangeofexp
8、ression,recentstudieshavedeterminedthatithascloseto1430presumedtargetmRNA,andtheseincludethep27,acellul
