中国组织工程研究

中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (9): 1390-1396.doi: 10.3969/j.issn.2095-4344.2017.09.015

• 干细胞移植 stem cell transplantation • 上一篇    下一篇

过表达miR-378骨髓间充质干细胞移植治疗心肌梗死

张秀敏1，2，于  波1，李学渊1   

  1. 1中国医科大学附属第一医院循环内科，辽宁省沈阳市  110001；2抚顺矿务局总医院循环内科，辽宁省抚顺市  113008
  • 出版日期:2017-03-28 发布日期:2017-03-31
  • 通讯作者: 于波，博士，教授，主任医师，中国医科大学附属第一医院循环内科，辽宁省沈阳市 110001
  • 作者简介:张秀敏，女，1979年生，辽宁省葫芦岛市人，满族，2003年中国医科大学毕业，副主任医师，主要从事心肌再生研究。
  • 基金资助:

    辽宁省科学技术计划项目(2012225084)；沈阳市科学技术项目(F12-193-9-03、F16-206-9-08)；辽宁省教育厅科学研究一般项目(L2013316)

The overexpression of miR-378 promotes the therapeutic effects of bone marrow mesenchymal stem cell transplantation on myocardial infarction

Zhang Xiu-min1, 2, Yu Bo1, Li Xue-yuan1   

  1. 1Department of Circulatory Medicine, the First Hospital of China Medical University, Shenyang 110003, Liaoning Province, China; 2Department of Circulatory Medicine, General Hospital of Fushun Mining Bureau, Fushun 113008, Liaoning Province, China
  • Online:2017-03-28 Published:2017-03-31
  • Contact: Yu Bo, M.D., Professor, Chief physician, Department of Circulatory Medicine, the First Hospital of China Medical University, Shenyang 110003, Liaoning Province, China
  • About author:Zhang Xiu-min, Associate chief physician, Department of Circulatory Medicine, the First Hospital of China Medical University, Shenyang 110003, Liaoning Province, China; Department of Circulatory Medicine, General Hospital of Fushun Mining Bureau, Fushun 113008, Liaoning Province, China
  • Supported by:

    the Science and Technology Plan of Liaoning Province, No. 2012225084; the Science and Technology Plan of Shenyang, No. F12-193-9-03, F16-206-9-08; the Scientific Research Project of Liaoning Provincial Department of Education, No. L2013316

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摘要：

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文题释义：
miRNAs：是长度为19-25个核苷酸的非编码小RNA分子，通过与靶基因mRNA 3’UTR互补配对，在转录后水平负性调控靶基因的表达，广泛参与细胞增殖、分化、凋亡、周期调控等生物学过程。
血管内皮生长因子、转化生长因子β1等细胞因子发挥心肌修复作用的机制：①抑制缺血区域心肌细胞凋亡，减小心肌梗死面积；②促进局部血管再生，增加心肌灌注；③激活心肌干细胞，增加梗死后心肌细胞再生。

 

摘要
背景：最近发现，miR-378能调节骨髓间充质干细胞的抗缺氧能力，减轻其在缺氧环境下的凋亡。
目的：观察miR-378过表达骨髓间充质干细胞在大鼠心肌梗死模型中的治疗作用。

方法：体外培养至第3代的大鼠骨髓间充质干细胞用反转录病毒感染建立miR-378过表达细胞系。选用成年SD大鼠，结扎前降支制作急性心肌梗死模型，随机分为3组：对照组(n=10)、BMSCnull组(n=16)、BMSCmiR-378组(n=16)。对照组仅注射50 μL生理盐水，后2组梗死局部注射50 μL生理盐水中含1×107个空病毒转染和miR-378模拟物转染的骨髓间充质干细胞。移植24 h后，利用TUNEL染色测定移植骨髓间充质干细胞的凋亡率，Western blotting检测移植区域内血管内皮生长因子、转化生长因子β1蛋白表达。移植4周后，利用心脏超声评价心功能，取局部心肌组织行组织学及分子生物学分析。
结果与结论：①移植24 h后，BMSCmiR-378组中凋亡细胞明显少于BMSCnull组(n=6，P < 0.001)，血管内皮生长因子、转化生长因子β1表达水平明显高于BMSCnull组(n=6，P < 0.001)；②移植4周后，BMSCmiR-378组中存活和分化为心肌细胞的骨髓间充质干细胞数量明显多于BMSCnull组(n=10，P < 0.001)。与其他2组相比，BMSCmiR-378组中新生血管数量明显增多(P < 0.001)；心肌梗死面积明显减小(P < 0.001)；左室射血分数明显增加(P < 0.05)；左室舒张末容积明显缩小(P < 0.05)。BMSCnull组中上述参数明显优于对照组(P < 0.05)；③结果表明，miR-378过表达可提高移植骨髓间充质干细胞的抗缺氧能力，继而促进心肌修复。

 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

ORCID: 0000-0002-2645-3900(张秀敏)

关键词: 干细胞, 移植, 心肌梗死, 骨髓间充质干细胞, miR-378, 血管新生, 心肌梗死面积, 左室射血分数

Abstract:

BACKGROUND: Recently, miR-378 has been shown to modulate the anti-hypoxia capacity of bone marrow mesenchymal stem cells (BMSCs) and reduce cell apoptosis under hypoxic conditions.

OBJECTIVE: To investigate the benefits of miR-378-upregulated BMSC transplantation in a rat model of acute myocardial infarction.
METHODS:Primary rat BMSCs were cultured in vitro. Until passage 3, the cells were infected with the lentivirus carrying synthetic miR-378 gene fragments. A rat model for acute myocardial infarction was constructed by ligating the left anterior descending artery. Thereafter, the animals were randomly assigned to three groups: control group (n=10), BMSCnull group (n=16) and BMSCmiR-378 group (n=16). In the latter two groups, 50 μL of normal saline containing 1×107 empty virus-transfected or miR-378-transfected BMSCs was injected into the region of myocardial infarction, respectively. Only 50 μL of normal saline was injected in the control group. Twenty-four hours later, the apoptosis of transplanted BMSCs was evaluated with TUNEL, and expression level of vascular endothelial growth factor and transforming growth factor-β was detected using western blot assay. Four weeks after treatment, the left ventricular function of rats was assessed by echocardiography, and then histological and molecular biology analyses were performed.

RESULTS AND CONCLUSION: At 24 hours postoperatively, there were less apoptotic BMSCs and higher expression levels of vascular endothelial growth factor and transforming growth factor-β in the BMSCmiR-378 group than in the BMSCnull group (n=6, P < 0.001). Four weeks later, there were more transplanted BMSCs and BMSCs-derived cardiomyocytes in the BMSCmiR-378 group than the BMSCnull group (n=10, P < 0.001). Moreover, increased new vessel density (P < 0.001), decreased infarcted area (P < 0.001), preserved left ventricular ejection fraction (P < 0.05), reduced left ventricular end-diastolic volume (P < 0.05) were found in the BMSCmiR-378 group, compared with the other two groups. The above parameters were better in the BMSCnull group than the control group (P < 0.05). Overall, the upregulation of miR-378 could enhance the capability of BMSCs against hypoxia, and consequently promote myocardial repair after implantation, providing a new strategy for cell therapy of myocardial infarction.

 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Myocardial Infarction, Bone Marrow, Mesenchymal Stem Cell Transplantation, MicroRNAs, Tissue Engineering

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