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1、重組人血管內(nèi)皮抑制素對(duì)血管內(nèi)皮細(xì)胞作用[摘要]目的:探討重組人血管內(nèi)皮抑制素(YH-16)對(duì)血管內(nèi)皮細(xì)胞(EC)的作用。方法:傳代培養(yǎng)ECV304,將不同濃度的YH-16作用于生長(zhǎng)旺盛的EC,以MTT法篩選有效濃度。以有效濃度的YH-16作用于EC,在不同時(shí)相點(diǎn)光鏡下觀察EC形態(tài),MTT檢測(cè)細(xì)胞活性,流式細(xì)胞術(shù)檢測(cè)細(xì)胞周期,實(shí)時(shí)熒光定量PCR檢測(cè)VEGF的表達(dá)量。結(jié)果:YH-16作用后,光鏡下可見(jiàn)EC發(fā)生彌散性壞死。MTT檢測(cè)顯示吸光度降低,流式細(xì)胞術(shù)示G2期細(xì)胞比例增多,實(shí)時(shí)熒光定量PCR檢測(cè)示VEGF表達(dá)量明顯降低。結(jié)論:YH-16可明顯抑制EC的生長(zhǎng)增殖
2、,可能通過(guò)抑制EC分泌VEGF,從而抑制EC增殖、遷移及血管的形成。[關(guān)鍵詞]重組人血管內(nèi)皮抑制素;血管內(nèi)皮細(xì)胞;細(xì)胞培養(yǎng)[中圖分類號(hào)]R96[文獻(xiàn)標(biāo)識(shí)碼]A[文章編號(hào)]1008-6455(2008)05-04EffectsofrecombinantedhumanangiostatinonvascularendothelialcellWUShang-min1,ZHAOYa-nan1,YANGLi2,WANGLu2,GUOShu-zhong2,LIUYin1,LIUDan2,LUKai-hua215(1.DepartmentofPlasticSurgery,Se
3、condAfiliatedHospital,KunmingMedicalCollege,Kunming650101,Yunnan,China.2.TheCentreofPlasticSurgery,theFourthMilitaryMedicalUniversity,Xi’an710032,Shaanxi,China)Abstract:ObjectiveToinvestigatethefunctionofrecombinantedhumanangiostatin(YH-16)onvascularendothelialcell(EC).Methods①Diffe
4、rentdosageofYH-16wasusedinthevascularendothelialcell,andtheeffectiveconcentration(10μg/ml)ofYH-16wasexaminedbyMTTmethod.②ThetestwasdividedintothreegroupsrandomlyasYH-16-experimentgroup,PBS-blankcontrolgroupandRPMI-1640medium-positivegroup.③Inthefollowing24h、48hand72hinterventions,th
5、emorphousofECwasobservedbyphotologyinvertedmicroscope,thecytoactivewasdetectedbymethodofMTTandthecellcyclewasinvestigatedbyflowcytometry(FLC).TheexpressionofVEGFwasdectectedbyreal-timefluorescentquantitatepolymerasechainreaction(Real-timePCR).ResultsECswerenecrosisintheeffectofYH-16
6、,15andmanycelldebriscanbeseenunderthelightmicroscope.TheabsorptancewasdegradedbyMTTassay(P<0.05),andFLCrevealedthepercentageofG2stagewasmult(P<0.05).TheexpressofVEGFwasobviouslydegraded(P<0.05).ConclusionYH-16cansuppresstheproliferationofvascularendothelialcellactivelyanditcansuppre
7、sstheDNAsynthesis.Simultaneously,itcaninhibitthesecretingofVEGF,andsuppresstheprolifetationofEC,alsomigrationandangiogenesis.Keywords:recombinanthumanvascularendostatin;vascularendothelialcell;cellculture血管畸形和血管瘤的病因目前仍不明了,治療上尚無(wú)針對(duì)性強(qiáng)的措施[1]。位于特殊部位如頭面部和生殖區(qū)的的病變常常出現(xiàn)毀容或影響功能等并發(fā)癥,甚至危及生命[2],因
8、此臨床上急需一種療效確切的治療手段。近