去铁胺促进血管内皮祖细胞靶向归巢和血管化的实验研究  

DFO Improves the Homing of EPCs and Neovascularization

作　　者：郑胜武[1] 黄雄梅 庄兢[1] 林根辉[1] 杨宇[1] 丁昕[1] ZHENG Sheng-wu;HUANG Xiong-mei;ZHUANG Jing;LIN Gen-hui;YANG Yu;DING Xin(Fujian Medical University Provincial College of Clinical Medicine(Department of Plastic Surgery,Fujian Provincial Hospital)Fuzhou 350001,Fujian,China)

机构地区：[1]福建医科大学省立临床医学院(福建省立医院整形外科),福建福州350001

出　　处：《中国美容医学》2018年第12期116-121,共6页Chinese Journal of Aesthetic Medicine

基　　金：福建省自然科学基金(2015J01427)

摘　　要：目的:探讨低氧模拟剂去铁胺(Desferrioxamine,DFO)模拟组织缺氧环境,促进血管内皮祖细胞(endothelial progenitorcells,EPCs)靶向归巢及促进血管化的可行性,并研究相关的信号通路。方法:分离、培养荧光素酶(luciferase)转基因Lewis大鼠的EPCs,作为移植细胞备用。实验动物NOD/SCID裸鼠被随机分为五组:对照组、EPCs、EPCsDFO、EPCs-DFO-AMD(AMD3100,趋化因子受体CXCR4抑制剂)、EPCs-DFO-LY(LY294002,磷酸肌醇3激酶(PI3K)抑制剂)组,每组10只。通过结扎裸鼠左侧股动脉造成左下肢缺血,经患侧股动脉移植EPCs,不同组别分别腹腔注射DFO(100mg/kg/d)、DFO和AMD3100(10mg/kg/d)、DFO和LY294002(20mg/kg/d),连续14d。分别从组织学,分子生物学,细胞学3个方面评价实验结果。结果:DFO的使用增加了裸鼠缺血下肢的血流灌注,改善了缺血下肢的功能,其新生毛细血管数量明显高于其他组,并且通过WesternBlot检测观察到缺血组织中VEGF(血管内皮生长因子),p-eNOS(磷酸化的一氧化氮合酶)和p-AKT(磷酸化蛋白激酶B)的分泌显著上调。在生物发光成像的研究中,DFO显著增加了EPCs向缺血患肢的迁移。这些作用均被PI3K抑制剂LY294002所抑制。结论:DFO的使用上调了EPCs向缺血患肢的迁移,改善了缺血患肢的血流灌注以及下肢功能,上调促血管生成因子的分泌,促进缺血组织的血管化,这些作用通过PI3K/AKT信号转导通路。Objective To investigate whether desferrioxamine(DFO)can improve the homing of endothelial progenitor cells(EPCs)and improve neovascularization in ischemic hindlimb of rats.The relevant mechanism was also explored.Methods EPCs of luciferase transgenic Lewis rats were isolated and cultured as transplantation cells.Experimental animal NOD/SCID nude mice were randomly divided into five groups of10:control,EPCs,EPCs-DFO,EPCs-DFO-AMD(AMD3100,CXCR4inhibitor),and EPCs-DFO-LY(LY294002,the PI3K inhibitor)groups.Left hindlimb ischemia was induced by ligating the left femoral artery in nude mice.EPCs were transplanted via the left femoral artery.After cell transplantation,intraperitoneal injection of DFO(100mg/kg/d),DFO and AMD3100(10mg/kg/d),DFO and LY294002(20mg/kg/d),were performed respectively to different groups for14days.The experimental results were evaluated from three aspects:histology,molecular biology and cytology.Results DFO treatment increased the perfusion of ischemic hindlimb and improved the function of ischemic hindlimb in nude mice.The number of new capillaries was significantly higher than that of other groups.Moreover,VEGF,p-eNOS and p-AKT were significantly increased through Western Blotdetection.In bioluminescence imaging,DFO significantly increased the migration of EPCs to ischemic limbs.All these effects were diminished by LY294002.Conclusion DFO treatment increased the migration of EPCs to ischemic limbs,improved blood perfusion and hindlimb function in ischemic limbs,up-regulated the secretion of pro-angiogenic factors and promoted neovascularization of ischemic tissue.These effects are mediated by the PI3K/AKT signal transduction pathway.

关 键 词：低氧模拟剂 低氧诱导因子 内皮祖细胞 归巢 血管化 PI3K/AKT信号通路 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]