CXCR4基因过表达HUCB-late EPCs移植对大鼠后肢缺血组织血管新生的影响  

Effect of revascularization on human umbilical cordblood-derived endothelial progenitor cells with CXCR4 over-expressed in hind limb ischemic tissue

作　　者：张百灵[1] 王照飞 张航[1] 何晋[1] 郑昭芬[1] 彭建强[1] 

机构地区：[1]湖南省人民医院心内科,湖南省长沙市410005 [2]长沙市第一人民医院心内科,湖南省长沙市410005

出　　处：《中国动脉硬化杂志》2017年第3期238-243,共6页Chinese Journal of Arteriosclerosis

基　　金：湖南省卫生厅科研基金课题(C2012-011);湖南省教育厅科研项目(15C0828)

摘　　要：目的观察转染rAAV_6-CXCR4和rAAV_6-GFP后,初步探索其是否对HUCB-late内皮组细胞(EPCs)的体外成血管能力及体内缺血组织血管新生有影响。方法用rAAV_6-CXCR4、rAAV_6-GFP感染HUCB-late EPCs,Western Blot检测CXCR4蛋白表达水平;应用基质胶成血管实验检测转染后细胞的体外成血管能力。构建SD大鼠后肢缺血模型,以一侧股动脉及分支结扎、离断的方法构建后肢缺血模型;18只大鼠建模成功后随机摸球法分为3组。空白组:损伤后6 h于局部注射500μL EGM-2;对照组:损伤后6 h于局部注射500μL含1×10~6个HUCB-late EPCs的EGM-2;实验组:损伤后6 h于局部注射500μL含1×10~6个CXCR4过表达HUCB-late EPCs的EGM-2。28天后,HE染色检测缺血后肢肌肉间、肌肉旁微小血管情况;免疫组织化学染色检测肌肉间、肌肉旁新生血管CD31阳性内皮细胞的表达情况,并计算CD31阳性新生微血管数量。结果 rAAV_6-CXCR4感染组HUCB-late EPCs中CXCR4蛋白的表达高于rAAV_6-GFP感染组和对照组。3组体外成血管能力差异无统计学意义(P>0.05)。体内实验,与空白组、对照组比较,实验组新生微血管表达的CD31^+内皮细胞明显增多(P<0.05)。结论 CXCR4过表达的HUCB-late EPCs可以促进SD大鼠后肢缺血组织新生微血管的形成。Ahn To evaluate the ability of capillary network formation of HUCB-late EPCs in vitro and in rat hind limb ischemic tissues, after transfected with rAAV-CXCR4 and rAAV-GFP. Methods HUCB-late EPCs were infected with rAAV6-CXCR4,rAAV6-GFP, and the expression of CXCR4 protein was detected by Western Blot analysis. In vitro,evaluate the ability of capillary network formation on Matrigel between the three groups. The mode of mouse hind limb ischemia was set up, by ligating the femoral artery and its branches. Following successful establishment, 18 rats were divided into three groups ran- domly： blank group were intramuscularly injected with 500 ILL EGM-2 at 6 hours following injury,and control group were intra- muscularly injected with 500 p^L EGM-2 at 6 hours following injury （contain 1~106 non-gene transfected HUCB-Iate EPCs）, and experiment group were intramuscularly injected with 500 ~xL EGM-2 at 6 hours following injury （ contain 1 x 106 CXCR4-gene transfected HUCB-late EPCs）. After 28 days, capillary network in hind limb muscles and next to muscles were detected by HE stain assay,and CD31 expressed in neovascularization endothelial cells were detected by immune histochemistry assay,and calcu- late the number of CD31 positive neovascularization, and calculate the number of CD31 positive neovascularization. Results Compared with infected rAAV6-GFP and non-infected group, the expression of CXCR4 protein was up-regulated after infected rAAV6-CXCRzl. In vitro, there is no significance of tube area in unit area among three types of HUCB-late EPCs （P〉0.05）.In the mode of mouse hind limb ischemia, compared with blank group and control group, CD31 positive cells of new capillary network in experimental group increased obviously （P〈0.05）. Conclusion CXCR4 over-expression of HUCBlate EPCs could promote neovascularization in rat hind limb ischemic tissues.

关 键 词：趋化因子受体4 后肢缺血模型 血管新生 

分 类 号：R363[医药卫生—病理学]