基质细胞衍生因子-1α与Bevacizumab在新生血管形成中的作用分析  

Role of SDF-1α and Bevacizumab in angiogenesis

作　　者：宋丹[1] 王陆飞[1] 车松天[1] 付金玲[1] 李旭 隋桂琴[1] 王桂云[1] 

机构地区：[1]吉林大学第二医院眼科,吉林长春130041

出　　处：《中国现代医学杂志》2011年第23期2855-2860,共6页China Journal of Modern Medicine

摘　　要：目的探讨基质细胞衍生因子-1α(stromalcellderivedfactor-1α,SDF-1α)与bevacizumab(Avastin)在新生血管形成中的作用及二者的关系。方法体外血管生成检测实验中人脐静脉内皮细胞(hu-manumbilicalveinendothelialcells,HUVECs)和人成纤维细胞共培养在24孔板中,对照组培养液仅含基础培养液,实验组在基础培养液中分别添加不同浓度重组人SDF-1α(10、100和1000ng/mL)、重组人VEGF165(10ng/mL)、Bevacizumab(10μg/mL)和/或抗SDF-1α抗体(10μg/mL),一共培养11d。然后固定、染色,测量各组形成毛细血管样结构的长度。细胞增殖检测采用5'-溴2'-脱氧尿嘧啶(BrdU)掺入法,用不同浓度重组人SDF-1α(10、100和1000ng/mL)、重组人VEGF165(100ng/mL)、Bevacizumab(100μg/mL)和/或抗SDF-1α抗体(10μg/mL)刺激HUVECs生长12h后用ELISA法进行BrdU检测。结果 SDF-1α显著促进了体外HUVECs毛细血管样结构形成和增殖,Bevacizumab(10μg/mL)明显抑制了VEGF165(10ng/mL)对HUVECs毛细血管样结构形成的促进了作用(P<0.01),却没有显著地抑制SDF-1α的促进作用,SDF-1α在Bevacizumab存在的情况下仍然显著地促进HUVECs毛细血管样结构的形成(P<0.05)。Bevacizumab(100μg/mL)明显抑制了VEGF165(100ng/mL)对HUVECs增殖的促进作用(P<0.05),却没有显著地抑制SDF-1α的促进作用,SDF-1α在Bevacizumab存在的情况下仍然显著地促进HUVECs增殖(P<0.05)。结论 SDF-1α在体外血管形成过程中起到了重要的促进作用。Bevacizumab不能抑制SDF-1α对新生血管形成的促进作用。【Objective】 To evaluate the role and relation of stromal cell-derived factor 1α（SDF-1α） and Bevacizumab（Avastin） in angiogenesis.【Methods】 For the capillary-like formation assay,an in vitro angiogenesis assay kit comprising co-cultures of human umbilical vascular endothelial cells（HUVECs） and human fibroblasts was used according to the manufacturer＇s instructions.Briefly,HUVECs and fibroblasts were co-cultured in 24-well plates with basal medium only（control） or basal medium containing various concentrations of recombinant human SDF-1α（10 ng/mL,100 ng/mL,1 000 ng/mL） and/or recombinant human VEGF165（10 ng/mL） and/or Bevacizumab（10 μg/mL） and/or anti-SDF-1α antibody（10 μg/mL）.HUVECs and fibroblasts were co-cultured for a total of 11 days.The cells were then fixed and stained.The total length of the capillary structures per group was measured.DNA synthesis measured by 5′-bromo-2′-deoxyuridine（BrdU） incorporation was performed as an index of cell proliferation.HUVECs were stimulated with various concentrations of recombinant human SDF-1α（10 ng/mL,100 ng/mL, 1 000 ng/mL） and/or recombinant human VEGF165（100 ng/mL） and/or Bevacizumab（100 μg/mL） and/or anti-SDF-1α antibody（10 μg/mL） for 12 hours.After the stimulation,BrdU ELISA chemiluminescence was analyzed according to the manufacturer＇s protocol.【Results】 SDF-1α significantly enhanced capillary-like formation and HUVECs proliferation.The activation of capillary-like formation by VEGF165（10 ng/mL） was effectively inhibited by Bevacizumab（10 μg/mL）（P〈0.01）.However,capillary-like formation promoted by SDF-1α was not inhibited by Bevacizumab.SDF-1α significantly enhanced capillary-like formation with Bevacizumab（P〈0.05）.The promotion of HUVECs proliferation by VEGF165（100 ng/mL） was significantly inhibited by Bevacizumab（100 μg/mL）（P〈0.05）.However,HUVECs proliferation promoted by SDF-1α was not inhibited by Bevacizumab.SDF-1α significantly enhanced

关 键 词：新生血管性青光眼 基质细胞衍生因子-1Α BEVACIZUMAB 血管生成 血管内皮生长因子 

分 类 号：R774.1[医药卫生—眼科] R775.9[医药卫生—临床医学]