中国组织工程研究

中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (36): 5325-5331.doi: 10.3969/j.issn.2095-4344.2016.36.001

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

高质量浓度肿瘤坏死因子α对骨髓间充质干细胞的损伤作用

李洁媚，王槐高，邓达师，卢  芳，张琛琛，刘国增，周艳芳   

  1. 广东医科大学病理生理学教研室，广东省东莞市  523808
  • 修回日期:2016-06-24 出版日期:2016-09-02 发布日期:2016-09-02
  • 通讯作者: 周艳芳，博士，广东医科大学病理生理学教研室，广东省东莞市 523808
  • 作者简介:李洁媚，女，1990年生，广东省梅县人，汉族，2016年广东医科大学毕业，硕士，主要从事神经病理生理学研究。 并列第一作者：王槐高，男，1978年生，湖南省武冈市人，汉族，2006年广东医科大学毕业，硕士，讲师，主要从事神经病理生理学研究。
  • 基金资助:

    广东省社会发展领域科技计划项目(2013B021800073)；东莞市科技局学科科研项目(2014108101049)

Tumor necrosis factor alpha-induced apoptosis in bone marrow mesenchymal stem cells

Li Jie-mei, Wang Huai-gao, Deng Da-shi, Lu Fang, Zhang Chen-chen, Liu Guo-zeng, Zhou Yan-fang   

  1. Department of Pathophysiology, Guangdong Medical University, Dongguan 525808, Guangdong Province, China
  • Revised:2016-06-24 Online:2016-09-02 Published:2016-09-02
  • Contact: Zhou Yan-fang, M.D., Department of Pathophysiology, Guangdong Medical University, Dongguan 525808, Guangdong Province, China
  • About author:Li Jie-mei, Master, Department of Pathophysiology, Guangdong Medical University, Dongguan 525808, Guangdong Province, China Wang Huai-gao, Master, Lecturer, Department of Pathophysiology, Guangdong Medical University, Dongguan 525808, Guangdong Province, China Li Jie-mei and Wang Hai-gao contributed equally to this work.
  • Supported by:

    the Scientific Research Plan for Social Development of Guangdong Province, No. 2013B021800073; the Scientific Research Project of Dongguan Science and Technology Department, No. 2014108101049

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摘要：

文章快速阅读：

 

文题释义：
原代培养：指直接从机体取细胞、组织和器官后立即进行培养，较为严格说是指成功传代之前的培养，此时的细胞保持原有细胞的基本性质，如果是正常细胞，仍然保留二倍体数。但实际操作上通常把第1 代至第10代以内的培养细胞统称为原代细胞培养。
肿瘤坏死因子α：可参与脑缺血后的损伤修复过程，调节脑缺血后血管发生和神经发生，在脑组织损伤恢复及神经保护方面具有重要意义。肿瘤坏死因子α对细胞的保护效应非常复杂，与其处理浓度、时间、实验模型、实验方法以及细胞来源息息相关。

 

摘要
背景：干细胞移植在脑缺血治疗中取得了良好效果，如何减少移植细胞的损伤成为治疗重点。
目的：探讨肿瘤坏死因子α对骨髓间充质干细胞的损伤作用及可能机制。
方法：原代培养SD大鼠骨髓间充质干细胞，高质量浓度肿瘤坏死因子α(200 μg/L)处理6 h，MTT比色法检测细胞存活率，Hoechst33342染色观测细胞凋亡，Annexin-V/PI双染检测细胞凋亡率，丙二醛、超氧化物歧化酶试剂盒检测细胞氧化应激水平，Western blot法检测Akt、p-Akt、FoxO1及p-FoxO1蛋白表达改变。
结果与结论：①相比对照组，高质量浓度肿瘤坏死因子α处理6 h后，骨髓间充质干细胞存活率下降，凋亡率上升；②肿瘤坏死因子α作用后，G1期和G2期细胞比例降低，S期细胞比例增高，氧化应激水平增高，p-Akt、p-FoxO1蛋白表达降低，差异有显著性意义(P < 0.05)；③结果提示，高质量浓度肿瘤坏死因子α可诱导骨髓间充质干细胞周期阻滞，诱导凋亡，提高其氧化应激水平，其损伤机制可能与抑制Akt-FoxO1信号通路有关。

 

 

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: 0000-0002-8477-6564(周艳芳)

关键词: 干细胞, 骨髓干细胞, 肿瘤坏死因子α, 骨髓间充质干细胞, Akt, FoxO1, 凋亡

Abstract:

BACKGROUND: Stem cell transplantation has achieved good results in the treatment of cerebral ischemia, and how to reduce apoptosis of transplanted cells has become the focus of the therapy.
OBJECTIVE: To investigate the injured effect of tumor necrosis factor alpha (TNF-α) on bone marrow mesenchymal stem cells and its mechanism.
METHODS: Primary cultured bone marrow mesenchymal stem cells from Sprague-Dawley rats were treated with 200 μg/L TNF-α for 6 hours. Cell vitality was assayed by MTT, and cell apoptosis was observed by Hoechst33342 staining. Apoptotic rate was detected by Annexin-V/PI double staining. Level of oxidative stress was evaluated by determination of malondialdehyde and superoxide dismutase levels. The protein expressions of phosphorylated-Akt, Akt, phosphorylated-FoxO1, FoxO1 were detected by western blot analysis.
RESULTS AND CONCLUSION: After treatment with TNF-α, the cell vitality of bone marrow mesenchymal stem cells decreased, the apoptotic rate increased, and the cells were arrested in the S phase. Moreover, the oxidative stress level was elevated, and the protein expression of phosphorylated-Akt and phosphorylated-FoxO1 was significantly reduced compared with the control group (P < 0.05). These results suggest that TNF-α at high level contributes to the S-stage arrest, responsible for the apoptosis processes of bone marrow mesenchymal stem cells via the Akt-FoxO1 pathway.

 

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: Tumor Necrosis Factor-alpha, Bone Marrow, Mesenchymal Stem Cells, Apoptosis, Tissue Engineering

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