資源描述:
《蛋白酶活化受體1與4在血小板活化中的作用機(jī)制》由會(huì)員上傳分享,免費(fèi)在線閱讀,更多相關(guān)內(nèi)容在行業(yè)資料-天天文庫。
1、中國實(shí)驗(yàn)血液學(xué)雜志JournalofExperimentalHematology2003;11(5):495-498·495·文章編號(hào)(ArticleID):1009-2137(2003)05-0495-04·論著·蛋白酶活化受體1與4在血小板活化中的作用機(jī)制11韓 悅,Jean2maxPasquet,AlanNurden,阮長耿1蘇州大學(xué)附屬第一醫(yī)院,江蘇省血液研究所,蘇州215006;UMR5533,CNRS,HopitalCardiologique,Pessac33604,France摘要 為了比較蛋白酶活化受體1和4(PAR1,PAR4)合成肽對血小板膜表面糖蛋白GPIbα表達(dá)及細(xì)胞
2、骨架的影響,揭示蛋白酶活化受體在血小板信號(hào)傳遞中的作用,選擇25μmol/LPAR1與250μmol/LPAR4兩類合成肽分別在不同時(shí)間段(0-60分鐘)活化血小板,應(yīng)用流式細(xì)胞儀測定血小板膜糖蛋白GPIbα及P2選擇蛋白的表達(dá),并結(jié)合SDS2PAGE與轉(zhuǎn)移電泳技術(shù)比較血小板活化前后細(xì)胞骨架GPIbα、肌動(dòng)蛋白、肌球蛋白的變化,同時(shí)對膜骨架成分進(jìn)行GPIbα免疫沉淀分析。結(jié)果顯示,PAR1與PAR4兩類合成肽均可促使血小板活化,其P2選擇蛋白水平顯著升高,GPIbα表達(dá)進(jìn)行性減少又出現(xiàn)逐漸回升的可逆性變化,各時(shí)間段引起的GPIbα改變在兩者之間都具有顯著差異(P<0.05),其中PAR1首先發(fā)
3、生作用,但PAR4維持時(shí)間更長。細(xì)胞骨架蛋白分析顯示肌動(dòng)蛋白與肌球蛋白協(xié)同GPIbα呈現(xiàn)先增加后減少的動(dòng)態(tài)變化。免疫印跡也表明與GPIbα相連的肌動(dòng)蛋白及肌球蛋白出現(xiàn)可逆性改變。結(jié)論:PAR1與PAR4在血小板信號(hào)傳遞過程中發(fā)揮了重要作用,它們能各自獨(dú)立地介導(dǎo)血小板活化,并導(dǎo)致GPIbα逆轉(zhuǎn),這與細(xì)胞骨架的積極參與相關(guān)。PAR1起效較快,PAR4維持作用更長久。關(guān)鍵詞 血小板活化;蛋白酶活化受體;糖蛋白Ibα;細(xì)胞骨架中圖分類號(hào) R3311143;R558文獻(xiàn)標(biāo)識(shí)碼 AMechanismofActionofProtease2ActivatedReceptors1and4inPlatele
4、tActivation11HANYue,Jean2MaxPasquet,AlanNurden,RUANChang2Geng1JiangsuInstituteofHematology,TheFirstAffiliatedHospitalofSuzhouUniversity,Suzhou215006,China;UMR553CNRS,HopitalCardiologique33604,Pessac,FranceAbstractThisstudywasdesignedtocomparetheeffectsofprotease2activatedreceptor1(PAR21)andprotease2
5、activatedreceptor4(PAR24)totheexpressionofplateletsurfaceGPIbαandcytoskeletonreorganization,thentoinvestigatetheroleofPARsinplateletsignaltransmission.PAR1(25μmol/L)andPAR4(250μmol/L)wereusedtostimulateplateletatdifferenttimepoints(0-60minutes),andtheplateletsurfaceGPIbα,actinandmyosinandP2selectinw
6、eredetectedwithflowcytometry,thealterationofGPIbα,actinandmyosinincytoskeletonwascomparedbyWesternblot,themembranecytoskeletonfollowedGPIbαimmunoprecipitationwasanalyzed.TheresultsshowedthatanincreaseofP2selectinandreversibledecreaseofGPIbαexpressionwereobtainedafterplateletactivationbyPAR1orPAR4,an
7、dadifferentkineticsofredistributionofGPIbαwasfoundforthetwopeptidesalloverthetimecourse(P<0105).PAR1actedmorepotentlyandrapidlythanPAR4,buttheeffectofPAR4persistedlongerinthecourseofplateletactivation