人脐带血CD34+细胞诱导小鼠缺血肢体血管生成  

Human Umbilical Cord Blood CD34+ Cells Induced Angiogenesis in Ischemic Limb of Mice

作　　者：陈作观 刁永鹏[1] 吴志远[1] 闫盛[1] 李拥军[1] CHEN Zuo-guan DIAO Yong-peng WU Zhi-yuan YAN Sheng LI Yong-jun(Department of Vascular Surgery, PUMC Hospital, CAMS and PUMC, Beijing 100730, China)

机构地区：[1]中国医学科学院北京协和医学院北京协和医院血管外科,北京100730

出　　处：《中国医学科学院学报》2016年第5期491-496,共6页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金(81270399)~~

摘　　要：目的观察体外增殖的人脐带血CD34+细胞对缺血肢体血管生成的影响,探讨CD34+细胞与血管生成的关系。方法采集人脐血细胞并进行CD34+细胞的提取、培养。建立小鼠左后肢缺血模型并随机分为CD34+细胞增殖组(n=5)、CD34+细胞组(n=5)和空白对照组(n=5)3组。采用Di I标记CD34+细胞示踪显像和抗人核抗原抗体(HNA)免疫组织化学方法评估人CD34+细胞向缺血区域的迁移能力,肢体温度、CD31免疫组织化学方法和转化生长因子β1(TGF-β1)m RNA表达评估缺血肢体血流改善情况。结果 Di I标记示踪及HNA显色可见扩增的CD34+细胞出现在小鼠缺血下肢的肌肉组织中并分布在血管周围。注射液体后第14天(t=5.421,P=0.001;t=0.616,P=0.000)和第28天(t=10.780,P=0.000;t=12.123,P=0.000),CD34+细胞增殖组和CD34+细胞组的温度改变百分比均明显高于对照组。注射液体后第28天,CD34+细胞增殖组和CD34+细胞组毛细血管密度分别为592.3±24.6(t=26.386,P=0.000)和530.7±25.5(t=21.502,P=0.000),均明显高于对照组的219.7±19.9;CD34+细胞增殖组和CD34+细胞组的TGF-β1mRNA表达量分别为(0.578±0.050)(t=12.376,P=0.000)和(0.504±0.080)copies(t=7.098,P=0.000),均明显高于对照组的(0.224±0.040)copies。结论体外培养的人脐带血CD34+细胞能向缺血区富集并诱导血管生成,促进血流复通,有望成为治疗下肢缺血的有效途径。Objective To observe the effect of the expanded human umbilical cord blood CD34 ＋ cells in ischemic limb of mice and analyse the relationship between the CD34 ＋ cells and angiogenesis.Methods Human umbilical cord blood was collected and CD34 ＋ cells were separated for expanding.Mice limbs ischemia models were established（ n = 15） and randomly divided into three groups：expanded CD34 ＋ cells group（ n =5）,fresh CD34 ＋ cells group（ n = 5）,and control group（ n = 5）.CD34 ＋ cells were detected by Di I dye tracing and antihuman nuclear antigen antibody（ HNA） immunohistochemical staining.The improvement of blood reperfusion was evaluated by indicators including limb temperature, CD31 staining, and transforming growth factor-β1（ TGF-β1） mRNA expression.Results On days 14（ t = 5.421,P = 0.001; t = 0.616,P =0.000） and 28（ t = 10.780,P = 0.000; t = 12.123,P = 0.000）,both expanded CD34 ＋ cells group and fresh CD34 ＋ cells group enjoyed better temperature improvement.Days 28 later,the vascular densities in the expanded CD34 ＋ cells group and the fresh CD34 ＋ cells group were 592.3 ± 24.6（ t = 26.386,P = 0.000） and530.7 ± 25.5（ t = 21.502,P = 0.000）,which were significantly higher than that in control group 219.7 ±19.9.The TGF-β1 mRNA expression in the expanded CD34 ＋ cells group and the fresh CD34 ＋ cells group were（ 0.578 ± 0.050） copies（ t = 12.376,P = 0.000） and（ 0.504 ± 0.080） copies（ t = 7.098,P = 0.000）,both over control group [（ 0.224 ± 0.040） copies].Conclusions In vitro culture of cord blood CD34 ＋ cells can emigrate to ischemic zone and induce angiogenesis to alleviate ischemia.Thus,it may provide a treatment option for lower limb ischemia.

关 键 词：血管生成 CD34+细胞 脐带血干细胞 肢体缺血 动物实验 

分 类 号：R654.4[医药卫生—外科学]