中国组织工程研究

中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (19): 2961-2967.doi: 10.3969/j.issn.2095-4344.2014.19.002

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

miR-378对缺血缺氧条件下骨髓间充质干细胞增殖及凋亡的影响

郭天柱，邢  越，侯婧瑛，周长青，钟婷婷，郑韶欣，王  彤   

  1. 中山大学孙逸仙纪念医院急诊科，广东省广州市  510120
  • 修回日期:2014-04-04 出版日期:2014-05-07 发布日期:2014-05-07
  • 通讯作者: 王彤，博士，博士生导师，教授，主任医师，研究员，中山大学孙逸仙纪念医院急诊科，广东省广州市 510120
  • 作者简介:郭天柱，男，1987年生，广西壮族自治区梧州市人，汉族，中山大学孙逸仙纪念医院在读硕士，主要从事急诊医学专业研究。
  • 基金资助:

    国家自然科学基金(81070125)“抗凋亡与促血管生成miRNA-378干预MSCs治疗心梗后心衰的机制研究”；国家自然科学基金(81270213)“ANGⅡ/AT1/SMAD/CX43通路在心肌干细胞提高心梗大鼠心电生理学稳定性和室颤阈值的作用机制研究”；广东省科技计划项目(2010B031600032)“抗凋亡与促血管生成miRNA-378干预MSCs治疗心梗后心衰的机制研究”；高校基本科研业务费中山大学青年教师重点培育项目(13ykzd16) “PPAR-γ/TGF-β1/Smad/CX43通路在PPAR-γ干预MSCs治疗心梗后心衰的疗效及机制研究”

miR-378 effects on the proliferation and apoptosis of bone marrow mesenchymal stem cells under hypoxic-ischemic condition

Guo Tian-zhu, Xing Yue, Hou Jing-ying, Zhou Chang-qing, Zhong Ting-ting, Zheng Shao-xin, Wang Tong   

  1. Department of Emergency, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, Guangdong Province, China
  • Revised:2014-04-04 Online:2014-05-07 Published:2014-05-07
  • Contact: Wang Tong, M.D., Professor, Chief physician, Investigator, Doctoral supervisor, Department of Emergency, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, Guangdong Province, China
  • About author:Guo Tian-zhu, Studying for master’s degree, Department of Emergency, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, Guangdong Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81070125, 81270213; the Scientific and Technological Plan of Guangdong Province, No. 2010B031600032; the Fundamental Research Funds for Universities-Key Cultivation Project of Young Teachers in Sun Yat-sen University, No. 13ykzd16

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465

被引次数

1

摘要：

背景：研究miR-378对缺血缺氧条件下骨髓间充质干细胞生长增殖、细胞凋亡等生物学行为的影响，为进一步改善骨髓间充质干细胞在梗死心肌中的生存提供新方法。
目的：观察miR-378转染后的骨髓间充质干细胞耐受缺血缺氧及促进血管形成的能力。
方法：体外培养的骨髓间充质干细胞分为未转染组和转染组，未转染组为未转染miR-378的骨髓间充质干细胞，转染组则采用化学合成的miR-378模拟物转染SD大鼠骨髓间充质干细胞；将两组骨髓间充质干细胞置于缺血缺氧(无血清，体积分数为1%O2、5%CO2，94%N2)环境中培养24 h后，应用锥虫蓝染色计数法、MTS法检测两组细胞在不同时间点的生长及增殖情况，TUNEL法检测细胞凋亡情况；将两组缺血缺氧后的培养基上清液分别刺激人脐静脉内皮细胞，观察血管形成情况。
结果与结论：缺血缺氧干预后48 h和72 h，转染组的骨髓间充质干细胞活细胞数明显高于未转染组(均为P < 0.01)；转染组的骨髓间充质干细胞表现出较高的增殖能力，缺血缺氧干预后24 h及48 h细胞增殖升高较未转染组明显，差异有显著性意义(P < 0.01，P < 0.05)；缺血缺氧后转染组骨髓间充质干细胞的凋亡比例明显下降(P < 0.05)。两组细胞均可促进血管腔样结构形成，但转染组血管管腔样结构较未转染组显著增多(P < 0.01)。研究结果提示miR-378可促进缺血缺氧后骨髓间充质干细胞的生长增殖并抑制其在缺血缺氧条件下的细胞凋亡，同时可提高其促血管生成的能力。


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


全文链接：

关键词: 干细胞, 骨髓干细胞, 骨髓间充质干细胞, miR-378, 缺血缺氧, 增殖, 凋亡, 血管形成能力, 国家自然科学基金

Abstract:

BACKGROUND: To investigate the influence of miR-378 on bone marrow mesenchymal stem cells proliferation and apoptosis under hypoxic-ischemic condition will provide a new method for further improving the survival of bone marrow mesenchymal stem cells in the infarcted myocardium.
OBJECTIVE: To observe the viability of bone marrow mesenchymal stem cells and their ability of angiogenesis under hypoxic-ischemic condition after miR-378 transfection.
METHODS: The bone marrow mesenchymal stem cells of Sprague-Dawley rat were cultured in vitro and divided into the non-transfection group and the transfection group. Mesenchymal stem cells were transfected with miR-378 mimic in the transfection group while the non-transfection group was taken as the control group. After transfection, the two groups were incubated under hypoxic-ischemic condition (serum-free medium,1% O2, 5% CO2, 94% N2) at 37 ℃ for 24 hours. Viability and proliferation of bone marrow mesenchymal stem cells were evaluated by the trypan-blue exclusion assay and MTS assay respectively. The cell apoptosis was detected by TUNEL assay. The culture supernatant of the two groups was collected separately and used as the conditioned medium after their exposure to hypoxic-ischemic environment. Endothelial cells were then co-cultured with the conditioned medium for the vasculature formation assay.
RESULTS AND CONCLUSION: After the experience of hypoxia-ischemia for 24 hours and 72 hours, there was a larger number of the living cells in the transfection group in contrast to the non-transfection group (both P < 0.01). Compared with the non-transfection group, miR-378-MSCs group presented a stronger ability of proliferation, and their proliferation rates were obviously higher at 24 and 48 hours (P < 0.01 and P < 0.05, respectively). The percentage of apoptotic cells in the transfection group was significantly declined under the hypoxic-ischemic condition (P < 0.05). The vasculature formation assay indicated that the lumen-like structures were found in both of the two groups, however, the number of lumen-like structures was remarkably increased in the transfection group (P < 0.01). miR-378 could promote the proliferation of bone marrow mesenchymal stem cells and suppress their apoptosis under hypoxic-ischemic condition. It could also enhance their ability of vasculogenesis.


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


全文链接：

Key words: bone marrow, mesenchymal stem cells, microRNAs, ischemia, anoxia, cell proliferation, apoptosis

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