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1、編號：刊用形式：文章編號：IL-8/CXCR2軸促進(jìn)兔內(nèi)皮祖細(xì)胞遷移、增殖及VEGF的表達(dá)毛梅1，2、呂學(xué)軍1、王藝1、徐劍鋮1，*（1第三軍醫(yī)大學(xué)新橋醫(yī)院呼吸病研究所重慶，400038；2廣州軍區(qū)武漢總醫(yī)院呼吸內(nèi)科武漢，430070）摘要目的觀察不同濃度白介素-8（interleukin-8，IL-8）對體外培養(yǎng)兔外周血內(nèi)皮祖細(xì)胞(endothelialprogenitorcells,EPCs)生物學(xué)功能的影響，探討趨化因子IL-8/CXCR2軸對EPCs血管生成功能的作用。方法密度梯度離心法獲取兔外周血單個核細(xì)
2、胞(mononuclearcells,MNCs)，接種至纖維連接蛋白(fibronectin,FN)包被的培養(yǎng)板上，培養(yǎng)7~10d后收集貼壁細(xì)胞鑒定EPCs，并用不同濃度IL-8的培養(yǎng)基繼續(xù)培養(yǎng)24h后采用MTT比色法、改良的Boyden小室實(shí)驗(yàn)，觀察EPCs的遷移和增殖能力；采用RT-PCR、Western-blot方法檢測CXCR2、VEGF的mRNA及蛋白表達(dá)水平。結(jié)果從兔外周血成功分離EPCs細(xì)胞，其增殖、遷移和VEGF的分泌受IL-8調(diào)節(jié)，且隨著IL-8濃度升高而增強(qiáng)。結(jié)論IL-8/CXCR2軸能顯著促進(jìn)
3、EPCs的遷移、增殖及VEGF的表達(dá)。關(guān)鍵詞趨化因子；白介素-8；內(nèi)皮祖細(xì)胞中圖法分類文獻(xiàn)標(biāo)識碼：ATheAxisofIL-8/CXCR2toPromoteProliferation,MigrationandVEGFExpressionofRabbits’EndothelialProgenitorCellsMAOMei1，2,LvXueJun1,Wangyi1,XUJian-cheng1，*(1,InstituteofHumanRespirationDiseases，XinqiaoHospital，ThirdMil
4、itaryMedicalUniversity．Chongqing400038;2,DepartmentofRespiratoryMedicine,WuhanGeneralHospitalofGuangzhouCommand,Wuhan430070)Abstract:ObjectiveToinvestigatetheproliferation,migrationandsecretionbiologicalfunctionofrabbits’peripheralbloodendothelialprogenitorcel
5、ls(EPCs)withdifferentconcentrationsofinterleukin-8.MethodsTotalmononuclearcells(MNCs)wereisolatedfromperipheralbloodbydensitygradientcentrifugation，andthenthecellswereplatedonfibronectin—coateculturedishes．Aftercultivationfor7days，attachedcellswereincubatedwit
6、hIL-8inaseriesofconcentrationsfor24h，thenattachedcellswerecharacterizedwithimmunofluorescenceandimmunohistochemistry．TheproliferationandmigrationactivitiesofEPCswereassayedwithMTTassayandmodifiedBoydenchamberassay，andreversetranscription(RT)一polymerasechainrea
7、ction(PCR)andwestern-blotwereusedtoassessmRNAandproteinexpressionsofCXCR2andVEGFofEPCs.ResultsTheEPCsfromrabbits’peripheralbloodweresuccessfullyisolatedandidentified．Theirproliferation,migrationandsecretionfunctionswereregulatedbyIL-8,andenhancedwithincreasing
8、concentrationofIL-8.ConclusionTheIL-8/CXCR2axiscansignificantlypromotetheEPCsproliferation,migration,andVEGFexpression.Keywords：chemotacticfactor；interleukin-8；Endothelialprogenito
