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1、DOI:10.16016/j.1000-5404.201502036金丝桃素光动力通过抑制HUVEC细胞增殖、迁移和管腔形成抑制血管生成张倩,李卓恒,李园园,李沉纹,管海燕,崔欢欢,李紫薇,卢来春(400042重庆,第三军医大学大坪医院野战外科研究所药剂科)[摘要]目的探讨金丝桃素光动力(hypericin-photodynamictherapy,HY-PDT)对人脐静脉内皮细胞(humanumbilicalveinendothelialcell,HUVEC)增殖、迁移及管腔形成的影响,研究抑制血管生成的作用机制。方法以血卟啉为阳性药,用金丝桃素及血卟
2、啉分别与HUVEC暗孵育24h后,给予585nm黄光照射(1.0J/cm2),24h后采用MTT法检测不同浓度HY对HUVEC细胞增殖的影响,Matrigel实验考察对HUVEC细胞管腔形成的作用,qRT-PCR检测Smad2基因mRNA的表达,Westernblot检测ERK1/2、p-ERK1/2、Smad2表达变化。结果HY-PDT能够抑制HUVEC细胞增殖、迁移及管腔形成;HY-PDT使HUVEC中ERK1/2磷酸化比率(p-ERK1/2/ERK1/2)显著降低(P<0.01),并显著下调Smad2mRNA及蛋白表达(P<0.01)。结论HY-
3、PDT通过抑制ERK1/2蛋白磷酸化活化、降低Smad2mRNA表达及蛋白水平,使HUVEC细胞增殖、迁移管腔形成受到抑制,抑制血管的生成。[关键词]金丝桃素;光动力;HUVEC;增殖;迁移;血管生成[中图法分类号][文献标志码]AHypericin-photodynamictherapyinhibitsangiogenesisthroughinhibitingHUVECcellproliferation,migrationandlumenformingZhangQian,LiZhuoheng,LiYuanyuan,LiChenwen,GuanHaiy
4、an,CuiHuanhuan,LiZiwei,LuLaichun(DepartmentofPharmacy,InstituteofSurgeryResearch,DapingHospital,ThirdMilitaryMedicalUniversity,Chongqing,400042,China)[Abstract]ObjectiveToinvestigatetheeffectofhypericin-photodynamictherapy(HY-PDT)onhumanumbilicalveinendothelialcell(HUVEC)prolife
5、rationandmigrationaswellaslumenformingandexplorethemechanismofHY-PDTinanti-angiogenesis.MethodsHematoporphyrin(HP)wasusedasthepositivecontrol.HUVECswereincubatedinmediacontainingHYorHPfor24hinthedark.Thecellswerethenirradiatedusinga585nmlightatadoseof1.0J/cm2.Afterthetreatment,t
6、hecellswereincubatedforanadditional24hinthedark.CellviabilitywasdeterminedbyMTTassay.TodetecttheeffectofHY-PDTonlumenforming,Matrigelexperimentwasimplemented.TotestthemRNAexpressionofSmad2andproteinlevelsofERK1/2,p-ERK1/2andSmad2,quantitativereal-timePCR(qRT-PCR)andWesternblotwe
7、recarriedout,respectively.ResultsHY-PDTinhibitedHUVECcellproliferation,migrationandlumenforming.Theratioofp-ERK1/2/ERK1/2wassignificantlyreducedinHUVECsfollowingHY-PDT(P<0.01).Additionally,HY-PDTremarkablydecreasedthemRNAandproteinexpressionlevel(P<0.01).ConclusionHY-PDTinhibite
8、dHUVECcellproliferationandmigrationaswellaslume