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1、重组人血管内皮抑制素对血管内皮细胞作用[摘要]目的：探讨重组人血管内皮抑制素(YH-16)对血管内皮细胞（EC）的作用。方法：传代培养ECV304，将不同浓度的YH-16作用于生长旺盛的EC，以MTT法筛选有效浓度。以有效浓度的YH-16作用于EC，在不同时相点光镜下观察EC形态，MTT检测细胞活性，流式细胞术检测细胞周期，实时荧光定量PCR检测VEGF的表达量。结果：YH-16作用后，光镜下可见EC发生弥散性坏死。MTT检测显示吸光度降低，流式细胞术示G2期细胞比例增多，实时荧光定量PCR检测示VEGF表达量明显降低。结论：YH-16可明显抑制EC的生长增殖
2、，可能通过抑制EC分泌VEGF，从而抑制EC增殖、迁移及血管的形成。[关键词]重组人血管内皮抑制素；血管内皮细胞；细胞培养[中图分类号]R96[文献标识码]A[文章编号]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，治疗上尚无针对性强的措施[1]。位于特殊部位如头面部和生殖区的的病变常常出现毁容或影响功能等并发症，甚至危及生命[2]，因
8、此临床上急需一种疗效确切的治疗手段。近
