中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (1): 66-71.doi: 10.12307/2022.984

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

miR-31促进骨髓间充质干细胞的增殖和迁移

张玉娟1，原一桐1，杜若琛1，田  峰1，付  媛2，王春芳1   

  1. 1山西医科大学实验动物中心，山西省太原市   030001；2山西医科大学基础医学院药理学教研室，山西省晋中市   030619
  • 收稿日期:2021-12-18 接受日期:2022-02-16 出版日期:2023-01-08 发布日期:2022-06-02
  • 通讯作者: 王春芳，博士，教授，山西医科大学实验动物中心，山西省太原市 030001
  • 作者简介:张玉娟，女，1994年生，山西省大同市人，汉族，山西医科大学在读硕士，主要从事干细胞与神经发育再生研究。
  • 基金资助:
    国家自然科学基金(81371384)，项目负责人：王春芳；国家自然科学青年基金(82001326)，项目负责人：原一桐；山西省应用基础研究计划项目(201901D211319)，项目负责人：原一桐

miR-31 promotes the proliferation and migration of bone marrow mesenchymal stem cells

Zhang Yujuan1, Yuan Yitong1, Du Ruochen1, Tian Feng1, Fu Yuan2, Wang Chunfang1   

  1. 1Experimental Animal Center, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China; 2Department of Pharmacology, College of Basic Medical Sciences, Shanxi Medical University, Jinzhong 030619, Shanxi Province, China
  • Received:2021-12-18 Accepted:2022-02-16 Online:2023-01-08 Published:2022-06-02
  • Contact: Wang Chunfang, MD, Professor, Experimental Animal Center, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China
  • About author:Zhang Yujuan, Master candidate, Experimental Animal Center, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China
  • Supported by:
    the National Natural Science Foundation of China, No. 81371384 (to WCF); the National Natural Science Youth Foundation of China, No. 82001326 (to YYT); the Applied Basic Research Program Project of Shanxi Province, No. 201901D211319 (to YYT)

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摘要：

文题释义：
细胞迁移：是突起延伸和胞体回缩通过特定的黏附点黏附到基质上的一个过程，主要依赖于细胞与细胞间的相互作用，是很多生理过程的重要核心。细胞迁移对发育、免疫防御和伤口愈合等各种生理过程至关重要。
miR-31：microRNA是一类内源性非编码的小分子RNA，可通过与靶基因特异性结合来抑制mRNA的翻译或稳定性。而miR-31是一种功能性的microRNA，在物种间高度保守，具有胚胎特异性并控制细胞命运和指导生理活动，也是细胞生物学的重要调节剂，参与各种细胞过程，包括细胞增殖和迁移。

背景：骨髓间充质干细胞已被广泛应用于临床治疗神经退行性相关疾病，但由于细胞在受损伤部位的存活率和迁移率低，从而降低了其疗效。
目的：探讨 miR-31是否能提高骨髓间充质干细胞的迁移和增殖能力。
方法：培养和鉴定C57BL/6小鼠骨髓间充质干细胞，将细胞分为对照组、miR-31 agomir组和miR-31 antagomir组。将第3代骨髓间充质干细胞接种于6孔板(1×105/孔)，待细胞达到50%-80%融合度，将miR-31 agomir和miR-31 antagomir用不含血清的DMEM/F12培养基稀释后添加到6孔板，转染24 h后，用CCK-8分析细胞的增殖水平以及Transwell实验分析细胞的迁移能力，Western blot检测基质金属蛋白酶2和CXC趋化因子受体4的蛋白表达。
结果与结论：①在不加转染试剂的情况下将miR-31成功转染至骨髓间充质干细胞，并发出红色荧光；②转染后，与对照组相比，
miR-31 agomir组细胞的增殖能力增强，与时间呈正比(P < 0.05)；与对照组相比，miR-31 agomir组细胞的迁移能力增强(P < 0.05)，基质金属蛋白酶2和CXC趋化因子受体4蛋白表达上调(P < 0.05)；③结果表明，miR-31可以提高骨髓间充质干细胞的增殖能力，并且促进骨髓间充质干细胞的迁移，这为间充质干细胞高效靶向迁移至损伤部位奠定了基础。

https://orcid.org/0000-0002-0633-0735 (张玉娟)

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

关键词: 间充质干细胞, 骨髓间充质干细胞, microRNA, miR-31, 细胞迁移, 脊髓损伤, CCXC趋化因子受体4, 基质金属蛋白酶2

Abstract: BACKGROUND: Bone marrow mesenchymal stem cells have been widely used in the clinical treatment of neurodegenerative-related diseases, but their efficacy is reduced by low cell survival and migration rates at the site of injury. 
OBJECTIVE: To investigate whether miR-31 can enhance the migration and proliferation of bone marrow mesenchymal stem cells. 
METHODS: Bone marrow mesenchymal stem cells from C57BL/6 mice were cultured and identified, and the cells were divided into control, miR-31 agomir, and miR-31 antagomir groups. Bone marrow mesenchymal stem cells at passage 3 were inoculated in six-well plates (1×105/well). When the cells reached 50%-80% fusion, miR-31 agomir and miR-31 antagomir were added to the six-well plates after dilution with serum-free DMEM/F12. After 24 hours of transfection, the proliferation level of cells was analyzed by CCK-8 assay as well as the migration ability of cells was analyzed by Transwell assay. Protein expression of matrix metalloproteinase 2 and CXC chemokine receptor 4 was detected by western blot assay. 
RESULTS AND CONCLUSION: (1) miR-31 was successfully transfected with bone marrow mesenchymal stem cells without transfection reagents and emitted red fluorescence. (2) After transfection, the proliferation ability of cells in the miR-31 agomir group was enhanced compared with the control group, which increased proportionally with time (P < 0.05). Compared with the control group, miR-31 promoted the migratory ability of bone marrow mesenchymal stem cells in the miR-31 agomir group (P < 0.05) and also upregulated protein expression of matrix metalloproteinase 2 and CXC chemokine receptor 4 (P < 0.05). (3) The results indicated that miR-31 could improve the proliferation ability of bone marrow mesenchymal stem cells and promote the migration of bone marrow mesenchymal stem cells, which provides a basic study for efficient targeting of mesenchymal stem cell migration to the site of injury. 

Key words: mesenchymal stem cell, bone marrow mesenchymal stem cell, microRNA, miR-31, cell migration, spinal cord injury, CXC chemokine receptor 4, matrix metalloproteinase 2

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