中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (10): 1808-1812.doi: 10.3969/j.issn.1673-8225.2010.10.020

• 干细胞与中医药 stem cells and traditional Chinese medicine • 上一篇    下一篇

Ca2+信号介导红景天苷促进小鼠骨髓间充质干细胞向神经细胞的定向分化

裴晶晶1，吴 润1，赵红斌2，刘旭东2，胡 军2，白孟海2   

  1. 1甘肃农业大学动物医学院，甘肃省兰州市  730070；
    2解放军兰州军区兰州总医院骨科研究所，甘肃省兰州市 730050
  • 出版日期:2010-03-05 发布日期:2010-03-05
  • 通讯作者: 赵红斌，博士，副教授，硕士生导师，解放军兰州军区兰州总医院骨科研究所，甘肃省兰州市 730050 zhao6703@ yahoo.com.cn
  • 作者简介:裴晶晶，女，1985年生，甘肃省兰州市人，汉族，甘肃农业大学在读硕士，主要从事干细胞基础方面的研究。 peijingjing0303@163.com
  • 基金资助:

    甘肃省自然科学基金资助项目（0710RJZA067），课题名称“组织诱导性人工角膜的构建”；
    兰州军区医药卫生科研基金（LXH-2007007），课题名称“组织诱导性神经导管的构建及对外周神经缺损修复的研究”。

Ca2+ signaling mediated salidrosides promotes directional differentiation of mouse bone marrow mesenchymal stem cells into nerve cells

Pei Jing-jing1, Wu Run1, Zhao Hong-bin2, Liu Xu-dong2, Hu Jun2, Bai Meng-hai2   

  1. 1College of Veterinary Medicine, Gansu Agricultural University, Lanzhou  730070, Gansu Province, China;
    2General Hospital of Lanzhou Military Area Command of Chinese PLA, Lanzhou  730050, Gansu Province, China
  • Online:2010-03-05 Published:2010-03-05
  • Contact: Zhao Hong-bin, Doctor, Associate professor, Master’s supervisor, General Hospital of Lanzhou Military Area Command of Chinese PLA, Lanzhou 730050, Gansu Province, China zhao6703@yahoo. com.cn
  • About author:Pei Jing-jing, Studying for master’s degree, College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, Gansu Province, China peijingjing0303@163.com
  • Supported by:

    the Natural Science Foundation of Gansu Province, No. 0710RJZA067*;
    the Research Foundation in Medical Science of Lanzhou Military Area Command, No. LXH-2007007*

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摘要：

背景：大量文献报道具有抗氧化作用的传统中药单体能促进骨髓间充质干细胞向神经细胞定向分化，红景天苷作为红景天的有效成分，具有较强的抗氧化功效。

目的：探讨红景天苷影响小鼠骨髓间充质干细胞向神经细胞定向分化的分子机制。

方法：体外培养的小鼠骨髓间充质干细胞达80%融合后，设立3组，空白对照组加入细胞完全培养液，诱导组、阳性对照组分别在其基础上加入20 mg/L红景天苷、0.1 mg/L神经生长因子，培养12 h，采用RT-PCR和Western blot法检测诱导后神经细胞相关基因和蛋白的表达。取诱导组细胞，分别加入含EGTA(细胞外Ca2+螯合剂)、Nifedipine(L型Ca2+通道阻断剂)、LY294002(IP3受体阻断剂)作用12 h，进行生物学信号传导通路检测。

结果与结论： ①诱导组可见神经元特异性烯醇化酶、β-Tubulin Ⅲ和Nurr1 mRNA的表达，阳性对照组上述基因的表达丰度均低于诱导组，空白对照组未见上述基因的表达；各组GFAP mRNA表达丰度均极低，诱导组c-fos mRNA呈高丰度表达。②与阳性对照组比较，诱导组能明显促进骨髓间充质干细胞神经元特异性烯醇化酶蛋白的表达；空白对照组无神经元特异性烯醇化酶蛋白的表达。③EGTA，Nifedipine，LY294002分别阻断细胞外Ca2+、L型Ca2+通道、IP3受体后，神经元特异性烯醇化酶和Nurr1基因、蛋白的表达均显著下调。结果证实红景天苷能使骨髓间充质干细胞定向分化为神经细胞，其生物学信号的传导与Ca2+信号有密切关系，IP3依赖的Ca2+信号途径是实现信号传导的重要方式之一。

关键词: 红景天苷, Ca2+信号, IP3, 神经元特异性烯醇化酶, 骨髓间充质干细胞

Abstract:

BACKGROUND: Tranditional Chinese medicine, which possesses anti-oxidation properties, can promote directional differentiation of mouse bone marrow mesenchymal stem cells (BMMSCs) into nerve cells. Salidrosides, as the effective constituent of Rhodiola, have strong anti-oxidation function.

OBJECTIVE: To investigate the molecule mechanism of salidrosides induced differentiation of mouse BMMSCs into nerve cells.

METHODS: When in vitro cultured BMMSCs reached 80% confluency, the cells were assigned into 3 groups. Cells in the control group were cultured by complete culture medium; those in the induction and positive control groups were cultured by complete culture medium adding 20 mg/L salidrosides or 0.1 mg/L nerve growth factors (NGF). The related gene and protein of nerve cells were detected using RT-PCR and Western blot method at 12 hours after culture. After that, the cells in the induction group were divided into 3 groups, the blocking agents EGTA (Ca2+ chelator), Nifedipine (L-type Ca2+ channel blocker) and LY294002 (IP3 receptor blocking pharmacon) were applied to block the cellular Ca2+ signal pathway respectively for 12 hours. RT-PCR and Western blot methods were used to study the signal transduction of the salidrosides.

RESULTS AND CONCLUSION: ①The expression of neuron specific enolase (NSE), β-Tubulin III, Nurr1 mRNA could be found in the induction and positive control groups, instead of the control group; The expression abundance of the positive control group was smaller than that of the induction group. The expression abundance of GFAP mRNA was very low in each group, but the c-fos mRNA was expressed abundantly in the induction group. ②Compared with the positive control group, the induction group could promote the NSE expression obviously, which was no expressed in the blank control group. ③The expression of NSE and Nurr1 were conspicuously down-regulated when the Ecto Ca2+ and L-type Ca2+ channel and IP3 receptor were blocked respectively. Salidrosides can induce the differentiation BMMSCs into nerve cells. Ca2+ signaling and IP3 dependent Ca2+ signaling pathway play an important role in transduction salidrosides signal in BMMSCs differentiation.

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