中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (14): 2523-2527.doi: 10.3969/j.issn.1673-8225.2010.14.011

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

高压氧对大鼠骨髓间充质干细胞分化和Wnt3表达的影响

谌崇峰，杨于嘉，姚  跃，王庆红，李  萌   

  1. 中南大学湘雅医院儿科，湖南省长沙市  410008
  • 出版日期:2010-04-02 发布日期:2010-04-02
  • 通讯作者: 杨于嘉，博士，教授，博士生导师，中南大学湘雅医院儿科，湖南省长沙市 410008 yyjcjcp@163.com
  • 作者简介:谌崇峰☆，女，1976年生，湖北省监利县人，汉族，中南大学湘雅医院在读博士，主要从事新生儿脑损伤方面的研究。 chenchfeng@yahoo.com.cn
  • 基金资助:

    国家自然科学基金资助项目(30672240)，课题名称“从Wnt信号途径探讨高压氧促缺氧缺血性脑损伤新生大鼠神经干细胞增殖分化机制”。

Effect of hyperbaric oxygen on differentiation and Wnt3 expression of bone marrow mesenchymal stem cells

Chen Chong-feng, Yang Yu-jia, Yao Yue, Wang Qing-hong, Li Meng   

  1. Department of Pediatrics, Xiangya Hospital, Central South University, Changsha   410008, Hunan Province, China
  • Online:2010-04-02 Published:2010-04-02
  • Contact: Yang Yu-jia, Doctor, Professor, Doctoral supervisor, Department of Pediatrics, Xiangya Hospital, Central South University, Changsha 410008, Hunan Province, China yyjcjcp@163.com
  • About author:Chen Chong-feng☆, Studying for doctorate, Department of Pediatrics, Xiangya Hospital, Central South University, Changsha 410008, Hunan Province, China chenchfeng@yahoo.com.cn
  • Supported by:

    the National Natural Science Foundation of China, No. 30672240*

PDF

358

被引次数

5

摘要：

背景：高压氧治疗可以促进缺氧缺血性脑损伤新生大鼠内源性神经干细胞的增殖和分化，且其机制与Wnt信号通路的活化有关，但在体外高压氧对干细胞分化的影响是否也通过Wnt信号通路起作用目前尚未见相关报道。
目的：观察高压氧对大鼠骨髓间充质干细胞分化的影响，认识其相关机制。
方法：全骨髓法分离培养大鼠骨髓间充质干细胞，贴壁纯化，取传至第3~5代细胞，加入含bFGF，EGF，B27的DMEM/F12培养基诱导培养24 h。诱导后细胞随机分为2组，对照组未做任何处理，高压氧组给予0.10 MPa的压力治疗，稳压时间60 min，稳压期间平均氧浓度不低于90%。免疫荧光染色检测巢蛋白、NSE，GFAP，O4等抗体的表达，Western-blot法检测Wnt3蛋白的表达。
结果与结论：骨髓间充质干细胞用神经干细胞经典培养基诱导后巢蛋白呈阳性表达。与对照组比较，高压氧组NSE，O4阳性细胞分化率均显著增加(P < 0.01)，GFAP阳性细胞分化率无明显差异(P > 0.05)，Wnt3蛋白的表达明显升高(P < 0.05)。高压氧可以促进骨髓间充质干细胞向神经元细胞和少突胶质细胞分化，但对星形胶质细胞的分化无明显影响，其机制可能与Wnt3蛋白的活化有关。

关键词: 高压氧, Wnt3, 诱导, 分化, 神经元, 少突胶质细胞, 骨髓间充质干细胞

Abstract:

BACKGROUND: Hyperbaric oxygen (HBO) treatment promotes the proliferation and differentiation of endogenous neural stem cells in neonatal rats following hypoxic/ischemic brain damage (HIBD). The Wnt signaling pathway is associated with neurogenesis. However, there are few data recording the role of HBO in the differentiation of neural stem cells in vitro.
OBJECTIVE: To observe the effect of HBO on differentiation and Wnt3 expression of bone marrow mesenchymal stem cells (BMSCs).
METHODS: BMSCs were isolated and cultured. The rat BMSCs of passages 3-5 were cultured in DMEM/F12 (1: 1) medium with basic fibroblast growth factor, epidermal growth factor and B27 for 24 hours. The induced BMSCs were randomly divided into two groups: control group (no treatment) and HBO group (HBO, 0.10 MPa, 60 minutes stabilizing pressure with at least 90% oxygen). The neuron specific encloase (NSE), glial fibrillary acidic protein (GFAP) and O4 marked oligodendrocyte immunocytochemistry were detected by immunofluorescent staining, and Wnt3 protein expression was detected by Western-blot.   
RESULTS AND CONCLUSION: BMSCs cultured in classic medium of neural stem cells could significantly induce the expression of nestin. The expression of NSE and O4 of HBO group was greater than control group (P < 0.01), but GFAP expression displayed no significant difference between the groups (P > 0.05). Western blot showed HBO could enhance the Wnt3 expression (P < 0.05). Results show that HBO can induce BMSCs to differentiate into neural cells and oligodendrocyte, which is correlated with the activation of the Wnt3 protein.

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