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任良强,侯晓晓,乔 平,王 圣,廖 旺,李 斌.脐血单个核细胞对急性心肌梗死大鼠血管新生的促进作用及机制研究[J].中国比较医学杂志,2021,31(5):72~77.
脐血单个核细胞对急性心肌梗死大鼠血管新生的促进作用及机制研究
Promotive effect and mechanism of umbilical cord blood mononuclear cells on angiogenesis in rats with acute myocardial infarction
投稿时间:2020-07-24  
DOI:10. 3969 / j.issn.1671-7856. 2021. 05. 012
中文关键词:  急性心肌梗死  脐血单个核细胞  血管新生
英文关键词:acute myocardial infarction  cord blood mononuclear cells  angiogenesis
基金项目:
作者单位E-mail
任良强 海南省人民医院心血管内科, 海口 570311 Shbai_7@ 163.com 
侯晓晓 海南省人民医院心血管内科, 海口 570311  
乔 平 海南省人民医院心血管内科, 海口 570311  
王 圣 海南省人民医院心血管内科, 海口 570311  
廖 旺 海南省人民医院心血管内科, 海口 570311  
李 斌 海南省人民医院心血管内科, 海口 570311  
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中文摘要:
       目的 探讨脐血单个核细胞(HCMNCs)对急性心肌梗死(AMI)大鼠血管新生的促进作用及机制。 方法 分离 HCMNCs 并用 BrdU 标记,选取 25 只大鼠采用冠状动脉结扎法建立急性心肌梗死大鼠模型,造模成功 20 只随机分为心梗组、HCMNCs 组,各 10 只,另取 10 只为假手术组。建模成功后,HCMNCs 组大鼠心肌梗死周围注入 HCMNCs,假手术组和模型组同部位注入 L-DMEM 培养基。移植 4 周后,超声心动图检测心功能,HE 染色观察心肌病理学变化,BrdU 染色检测移植细胞存活情况,免疫组化染色检测心肌梗死边缘微血管密度( MVD)、 CD31、血管内皮生长因子(VEGF)蛋白表达,Western blot 法检测心肌组织中 CD31、VEGF 蛋白水平。 结果 与假手术组比较,心梗组和 HCMNCs 组左室收缩末期内径(LVEDs)、左室舒张末期内径(LVEDd)水平升高,左室射血分数 (LVEF)、左室短轴缩短率(FS)水平降低(P<0. 05);与心梗组比较,HCMNCs 组 LVEDs、LVEDd 水平降低,LVEF、FS 水平升高(P<0. 05)。 HE 染色显示,假手术组心肌细胞结构正常,心梗组心肌细胞排列紊乱,有大量炎性细胞浸润;HCMNCs 组心肌细胞结构基本正常。 BrdU 染色显示,假手术组和心梗组均未检测到 BrdU 标记的阳性细胞, HCMNCs 组梗死区可见散在分布的 BrdU 阳性细胞,参与血管壁的组成。与假手术组比较,心梗组和 HCMNCs 组 CD31、VEGF 蛋白光密值及蛋白相对表达量和 MVD 减少(P<0. 05);与心梗组比较,HCMNCs 组 CD31、VEGF 蛋白光密值及蛋白相对表达量和 MVD 增加(P<0. 05)。 结论 HCMNCs 能促进 AMI 大鼠侧支循环建立,诱导血管新生,对缺血性心功能有明显改善作用。
英文摘要:
       Objective To explore the promotive effect and mechanism of human cord blood mononuclear cells (HCMNCs) on angiogenesis in rats with acute myocardial infarction ( AMI). Methods HCMNCs were isolated and labeled with BrdU. Twenty rats were selected to establish the acute myocardial infarction model by coronary artery ligation. The rats were randomly divided into myocardial infarction and HCMNC groups with 10 rats in each group. Another 10 rats were used as the sham operation group. After successful model establishment, HCMNCs were injected around the myocardial infarction in the HCMNC group and L-DMEM was injected into the same site of sham operation and model groups. Four weeks after transplantation, echocardiography was used to assess cardiac functions, HE staining was used to observe myocardial pathological changes, BrdU staining was used to detect transplanted cell survival, and immunohistochemical staining was used to analyze the microvessel density (MVD) and protein expression of CD31 vascular endothelial growth factor (VEGF). Western blott was used to measure CD31 and VEGF protein levels in myocardial tissue. Results Compared with the sham operation group, the left ventricular end systolic dimension ( LVEDs ) and left ventricular end diastolic dimension (LVEDd) were increased and the left ventricular ejection fraction (LVEF) and fraction shortening (FS) were decreased in myocardial infarction and HCMNC groups (P<0. 05). Compared with the myocardial infarction group, LVEDs and LVEDd were decreased, and LVEF and FS were increased in the HCMNC group (P<0. 05). HE staining showed that the structure of myocardial cells was normal in the sham operation group and the arrangement of myocardial cells was disordered in the myocardial infarction group with a large amount of inflammatory cell infiltration. The structure of myocardial cells in the HCMNC group was essentially normal. BrdU-positive cells were not detected in sham operation or myocardial infarction groups and scattered BrdU-positive cells were seen in the infarct area of the HCMNCs group, which were involved in formation of the blood vessel wall. Compared with the sham operation group, relative expression of CD31 and VEGF proteins and MVD were decreased in myocardial infarction and HCMNC groups (P<0. 05), and compared with the myocardial infarction group, relative expression of CD31 and VEGF proteins and MVD were increased (P< 0. 05). Conclusions HCMNCs promote the establishment of collateral circulation in AMI rats, induce angiogenesis, and significantly improve ischemic heart functions.
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