中国组织工程研究

中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (41): 6573-6577.doi: 10.3969/j.issn.2095-4344.2014.41.003

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

缺氧环境中胸腺素β4基因转染骨髓间充质干细胞凋亡率及Bax和Bcl-2的表达

刘德胜1，肖诗亮2，王 博2，周 诚2   

  1. 1宜宾市第二人民医院胸心外科，四川省宜宾市 644000；2华中科技大学同济医学院附属协和医院心外科，湖北省武汉市  430022
  • 修回日期:2014-07-25 出版日期:2014-10-01 发布日期:2014-10-01
  • 通讯作者: 周诚，博士，主治医师，华中科技大学同济医学院附属协和医院心外科，湖北省武汉市 430022
  • 作者简介:刘德胜，男，1985年生，重庆市人，汉族，2012年华中科技大学毕业，硕士，医师，主要从事冠心病临床与基础研究。
  • 基金资助:

    武汉晨光计划项目(200950431177)

Expressions of Bcl-2 and Bax and apoptotic rate of bone marrow mesenchymal stem cells transfected with thymosin beta4 in a hypoxic environment

Liu De-sheng1, Xiao Shi-liang2, Wang Bo2, Zhou Cheng2   

  1. 1Department of Cardiothoracic Surgery, Second People’s Hospital of Yibin City, Yibin 644000, Sichuan Province, China; 2Department of Cardiac Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China
  • Revised:2014-07-25 Online:2014-10-01 Published:2014-10-01
  • Contact: Zhou Cheng, M.D., Attending physician, Department of Cardiac Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China
  • About author:Liu De-sheng, Master, Physician, Department of Cardiothoracic Surgery, Second People’s Hospital of Yibin City, Yibin 644000, Sichuan Province, China
  • Supported by:

    Morning Light Plan of Wuhan City, No. 200950431177

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摘要：

背景：研究表明体外培养细胞加入胸腺素β4能增加细胞的抗凋亡能力，若在缺氧环境中上调胸腺素β4基因在骨髓间充干细胞中的表达，其抗凋亡能力如何改变，目前鲜见报道。

目的：观察胸腺素β4基因修饰的骨髓间充质干细胞在缺氧环境中的凋亡率改变，并探讨其是否通过调控Bax、Bcl-2表达影响凋亡能力。
方法：将携带有胸腺素β4基因的慢病毒转染骨髓间充质干细胞，转染完毕后利用Western blot法检测胸腺素β4在骨髓间充质干细胞中的表达。将细胞分为胸腺素β4转染组、对照病毒组、未转染组，分别置于缺氧环境中。流式细胞仪检测3组细胞凋亡率；Western blot法检测3组细胞中Bax和Bcl-2蛋白表达情况。
结果与结论：Western blot检测结果示，胸腺素β4基因在骨髓间充质干细胞中成功表达。流式细胞仪结果示，胸腺素β4转染组细胞凋亡率较对照病毒组、未转染组低，而对照病毒组、未转染组凋亡率差异无显著性意义。Western blot结果显示，Bcl-2蛋白在胸腺素β4转染组中表达量较对照病毒组、未转染组高，Bax蛋白在胸腺素β4转染组表达量较对照病毒组、未转染组低，而对照病毒组、未转染组中Bax、Bcl-2表达差异无显著性意义。提示过表达胸腺素β4基因可增加骨髓间充质干细胞在缺氧环境中的抗凋亡能力，其可能的作用机制是调控Bax和Bcl-2蛋白表达水平。

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, 胸腺素β4, 骨髓间充质干细胞, 基因, 凋亡, Bax, Bcl-2

Abstract:

BACKGROUND: Studies have shown that thymosin β4 can improve the anti-apoptotic ability of a variety of cells, but the reports describing the changes in the anti-apoptotic ability of bone mesenchymal stem cells modified with thymosin β4 gene in the hypoxic environment are rare.

OBJECTIVE: To observe the change in the apoptotic rate of bone mesenchymal stem cells modified with thymosin β4 gene in the hypoxic environment, and to explore whether thymosin β4 affects the apoptosis of bone marrow mesenchymal stem cells via regulating the expression of Bax and Bcl-2.
METHODS: Bone marrow mesenchymal stem cells were transfected with the recombinant lentiviral vector over-expressing the thymosin β4 gene, and then we observed the expression of thymosin β4 using western blot assay. Cells were divided into three groups: thymosin β4 transfection group, control virus group, and untreated group. All three groups were placed in a hypoxic environment. The apoptotic rate of bone marrow mesenchymal stem cells was evaluated by the flow cytometry assay. The expression of Bax and Bcl-2 protein was detected by western blot.
RESULTS AND CONCLUSION: Thymosin β4 gene was expressed successfully in the bone marrow mesenchymal stem cells showed by the western blot. The apoptotic rate of bone marrow mesenchymal stem cells in the hypoxic environment was lower in the thymosin β4 transfection group than the control virus group and untreated group; while there was no difference between the latter two groups. Western blot results showed that the expression of Bcl-2 protein was higher in the thymosin β4 transfection group than the control virus group and untreated group, and the expression of Bax protein was lower in the thymosin β4 transfection group than the control virus group and untreated group. No difference in the expression of Bax and Bcl-2 was found between the control virus group and untreated group. These findings indicate that thymosin β4 overexpression can improve the anti-apoptotic ability of bone mesenchymal stem cells modified in the hypoxic environment, and its possible mechanism is through regulating the expression of Bax and Bcl-2 protein.

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: stem cells, mesenchymal stem cells, thymosin, apoptosis

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