去铁胺促进BMSCs靶向归巢和血管新生的实验研究  

Experimental study on the effect of desferrioxamine on targeted homing and angiogenesis of bone marrow mesenchymal stem cells

作　　者：郑胜武[1] 杜子婧[2] 黄雄梅 庄兢[1] 林根辉[1] 杨宇[1] 丁昕[1] 昝涛[2] ZHENG Shengwu;DU Zijing;HUANG Xiongmei;ZHUANG Jing;LIN Genhui;YANG Yu;DING Xin;ZAN Tao(Department of Plastic Surgery,Fujian Provincial Hospital,Fujian Medical University,Fuzhou Fujian,350001,P.R.China;Department of Plastic Surgery,Shanghai Ninth People's Hospital,Shanghai JiaoTong University School of Medicine,Shanghai,200011,P.R.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,福州350001 [2]上海交通大学医学院附属第九人民医院整复外科,上海200011

出　　处：《中国修复重建外科杂志》2019年第1期85-92,共8页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金资助项目(81471857);福建自然科学基金资助项目(2015J01427)~~

摘　　要：目的应用低氧模拟剂去铁胺(desferrioxamine,DFO)模拟组织缺氧环境,观察其是否能促进BMSCs在大鼠随意皮瓣中的归巢和血管新生。方法分离、培养荧光素酶转基因Lewis大鼠的BMSCs和成纤维细胞(fibroblast,FB)。选用4周龄Lewis雄性大鼠40只,在其背部形成10 cm×3 cm大小矩形皮瓣,然后随机分成4组,每组10只:A组于大鼠球后静脉丛注射200μL PBS;B、C组同上法分别注射浓度为1×10~6个/mL的FB和BMSCs 200μL;D组同C组方法注射BMSCs后,腹腔注射DFO[100 mg/(kg·d)],连续7 d。术后7 d,观察各组大鼠皮瓣成活情况并计算皮瓣成活率,采用激光散斑血流成像仪检测皮瓣血流情况;术后30 min及1、4、7、14 d采用生物发光成像检测移植细胞在大鼠体内分布情况;术后7 d行CD31免疫荧光染色计算毛细血管密度,免疫荧光检测基质细胞衍生因子1(stromal cell derived factor 1,SDF-1)、EGF、FGF及Ki67的表达情况;利用荧光素酶抗体标记移植的BMSCs,免疫荧光染色观察其是否参与损伤组织修复。结果术后7 d各组缺血皮瓣坏死边界已明确,C、D组皮瓣成活率明显高于A、B组,D组高于C组(P<0.05)。激光散斑血流成像仪检测示,C、D组皮瓣血流值显著高于A、B组,D组高于C组(P<0.05)。生物发光成像示BMSCs随时间变化逐渐向缺血缺氧区迁移,最终分布到缺血组织中;术后14 d D组的光子信号明显强于其他组(P<0.05)。CD31免疫荧光染色示,C、D组毛细血管密度显著高于A、B组,D组高于C组(P<0.05)。C、D组SDF-1、EGF、FGF及Ki67的表达明显强于A、B组,D组强于C组。术后7 d移植的荧光素酶标记BMSCs表达于组织的动脉弹力层、毛细血管处和毛囊处。结论 DFO可以加速BMSCs向随意皮瓣缺氧区域的迁移归巢,加速BMSCs在缺血组织中的分化,同时促进缺血组织血管新生。Objective To investigate whether desferrioxamine (DFO) can enhance the homing of bone marrow mesenchymal stem cells (BMSCs) and improve neovascularization in random flaps of rats. Methods BMSCs and fibroblasts (FB) of luciferase transgenic Lewis rats were isolated and cultured. Forty 4-week-old Lewis male rats were used to form a 10 cm×3 cm rectangular flap on their back. The experimental animals were randomly divided into 4 groups with 10 rats in each group: in group A, 200 μL PBS were injected through retrobulbar venous plexus; in group B, 200 μL FB with a concentration of 1×106 cells/mL were injected; in group C, 200 μL BMSCs with a concentration of 1×10^6 cells/mL were injected; in group D, cells transplantation was the same as that in group C, after cells transplantation, DFO [100 mg/(kg·d)] were injected intraperitoneally for 7 days. On the 7th day after operation, the survival rate of flaps in each group was observed and calculated; the blood perfusion was observed by laser speckle imaging. Bioluminescence imaging was used to detect the distribution of transplanted cells in rats at 30 minutes and 1, 4, 7, and 14 days after operation. Immunofluorescence staining was performed at 7 days after operation to observe CD31 staining and count capillary density under 200-fold visual field and to detect the expressions of stromal cell derived factor 1 (SDF-1), epidermal growth factor (EGF), fibroblast growth factor (FGF), and Ki67. Transplanted BMSCs were labeled with luciferase antibody and observed by immunofluorescence staining whether they participated in the repair of injured tissues. Results The necrosis boundary of ischemic flaps in each group was clear at 7 days after operation. The survival rate of flaps in groups C and D was significantly higher than that in groups A and B, and in group D than in group C (P<0.05). Laser speckle imaging showed that the blood perfusion units of flaps in groups C and D was significantly higher than that in groups A and B, and in group D than in group C (P<0.05). Biolum

关 键 词：低氧模拟剂 去铁胺 BMSCS 归巢 血管新生 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]