IntJOphthalmol，Vo1．8，No．5，May2008www．IJO．caTel：029-8224517283085628EmaU：IJO．2000@163．COITI·实验论著·黄酮对体外培养人角膜内皮细胞的影响及其抗氧化作用李凌，樊廷俊，杨秀霞，丛日山，赵君，王晶基金项目：中罔国家“十五”863项目资助(No．2001AA625050)；国家“十一五”863项目资助(No．2006AA02A132)作者单位：(266003)中同山东省青岛市，中同海洋大学海洋生命学院海洋生物系；(266071)中国山东省青岛市，中国科学院海洋研究所实验海洋生物学开放实验室作者简介：李凌，女，理学硕十，在读博士。通讯作者：樊廷俊，男，教授，理学博士，中国海洋大学生命学院副院长、国家生命科学与技术人才培养基地主任、国家海洋生命科学实验教学示范中心主任，研究方向：动物细胞工程与细胞分化，主要成果：在国际上建立了首个未经任何肿瘤病毒或癌基因转染的家免和人角膜内皮细胞系等．tjfan@mail．OUC．edu．cn收稿日期：2008-04-06修回日期：2008-23E仟ectsandanti0xidativeactivitiesoffla．voneonhumancorneaIend0the¨aIcells／nvitroLingLi一，Ting-JunFan，Xiu-XiaYang，Ri-ShanCong，JunZhao，JingWangFoundationiterns：National863Programduringthe10thFive—YearPlanPeriod(No，2o01AA625050)：National863Programduringthe11thFive—YearPlanPeriodfNo．2OHD6AA02A132)DepartmentofMarineBiology，FacuhyofManneLifeScience，OceanUniversityofChina，Qingdao266003，ShandongProvince，China；LaboratoryofExperimentalMafineBiology．InstituteofOceanology，ChineseAcademyofSciences，Qingdao266071，ShandongProvince，ChinaC0rresp0ndenceto：Ting—JunFan，DepartmentofMarineBiology，FacultyofMarineLifeScience，OceanUniversityofChina．Qingdao266003．ShandongProvince．China．tifan@mail，oue．edu．cnReceived：20084_o6Accepted：2OHD8—O4_23Abstract·AlM：T0evaluatetheefectandantioxidativeactivitiesofflavoneonhumancornealendothelialcellsfHCEC)invitro．·METHODS：Di仟erentcOncentratiOnsofflavonewereaddedintotheculturemediumofHCEC．andthemorphologyofHCECwasobservedandtheirapoptosiswasverifiedbyacridineorange／ethidiumbromide(A0／EB)doublefluorescentstainingandDNAagarosegelelectrODhOresjs．Afterdi仟erentcOncentratiOnsofflavonewereaddedintotheculturemediumofHCECandculturedfor30minutes，600vmol／Lhydrogenperoxide(H2O2)wasfurtheraddedintotheculturemediumofHCEC．andthemorphologyofHCECwasobservedandtheirapoptosiswasverifiedbvAO／EBdoublefluorescentstainingandDNAagarosegelelectrODhOresis．·RESULTS：FlavonehadnoinjuryefectsonHCECatlowcOncentratiOns(1owerthan70vmol／L)．wh_lehadobviousapoptosis—inducingefectstoHCECathighcOncentrat_0ns(higherthan85vmol／L)．Preincubationwith55IJmol／Land70pmol／Lflavonecouldinh_bitsignificantlytheapoptosisofHCECinducedbV600IJmol／LH，O，，whilepreincubationwith85vmol／Land100vmol／Lflavonecouldnot．·CONCLUSlON：Flavonehasprominentantioxidativeacti—vitiesonHCECfrOmoxidantinducedapoptosisatconcen—trationof55—70IJmol／L，buthasobviousapoptosis—inducingefectsonHCECatconcentrationhigherthan85IJmol／L．·KEWVORDS：humancornealendothelialcells；apoptosis；hydrogenperoxide；flavone；antioxidativeactivities¨L，FanTJ，YangXX，eta1．Effectsandantioxidativeactivitiesofflavoneonhumancornealendothelialcellsinvitro．IntJOphthalmoluodYankeZazbi2008；8(5)：881—885摘要目的：确定黄酮对人角膜内皮细胞(humancornealendothe—lialcells，HCEC)的影响及其对因氧化损伤所致细胞凋亡的抗氧化保护作用。方法：利用体外培养的HCEC，通过添加不同浓度的黄酮继续培养，或经不同浓度黄酮预处理后再添加过氧化氢(H，O，)继续培养，采用光镜形态观察、吖啶橙／溴化乙锭(AO／EB)双染色和DNA琼脂糖...
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