中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (45): 8520-8523.doi: 10.3969/j.issn.2095-4344.2012.45.030

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

杜仲醇提取物诱导骨髓间充质干细胞成骨分化中的Wnt信号途径

张 贤1，朱丽华1，钱晓伟2，谭湘陵2   

  1. 1南京中医药大学无锡附属医院，江苏省无锡市 214000
    2南通大学生命科学学院，江苏省南通市 226019
  • 收稿日期:2012-01-09 修回日期:2012-02-28 出版日期:2012-11-04 发布日期:2012-11-04
  • 通讯作者: 谭湘陵，硕士，教授，南通大学生命科学学院，江苏省南通市 226019 tanxl@ntu.edu.cn
  • 作者简介:张贤，男，1971年生，湖南省永州市人，汉族，1994年湖南中医药大学毕业，副主任医师，主要从事脊柱脊髓损伤的基础与临床研究。 zhangxian0772@sina.com

Wnt signaling pathway for osteogenic differentiation of bone marrow mesenchymal stem cells induced by eucommia bark

Zhang Xian1, Zhu Li-hua1, Qian Xiao-wei2, Tan Xiang-ling2   

  1. 1Wuxi Affiliated Hospital of Nanjing University of Traditional Chinese Medicine, Wuxi 214000, Jiangsu Province, China
    2College of Life Sciences, Nantong University, Nantong 226019, Jiangsu Province, China
  • Received:2012-01-09 Revised:2012-02-28 Online:2012-11-04 Published:2012-11-04
  • Contact: Tan Xiang-ling, Master, Professor, College of Life Sciences, Nantong University, Nantong 226019, Jiangsu Province, China tanxl@ntu.edu.cn
  • About author:Zhang Xian, Associate chief physician, Wuxi Affiliated Hospital of Nanjing University of Traditional Chinese Medicine, Wuxi 214000, Jiangsu Province, China zhangxian0772@sina.com

PDF

528

被引次数

22

摘要：

背景：近年来，中药及中药有效部分对骨质疏松的干预和治疗作用的报道较多，但涉及细胞成骨分化调控的信号途径的报道较少。
目的：观察杜仲诱导大鼠骨髓间充质干细胞成骨分化过程中Wnt信号途径相关基因表达的变化。
方法：将第3代SD大鼠骨髓间充质干细胞，接种到6孔培养板中，每孔1×103个细胞，24 h后更换诱导培养基(含体积分数为7.5%胎牛血清的DMEM/F12(1∶1)加1/1 000浓度的杜仲醇提取物)。阴性对照组仍为正常培养基培养。诱导8 h，1 d，3 d和7 d时采用RT-qPCR法测定Wnt信号途径中Fzd和LRP受体系列、β-catenin、核内Wnt调控靶基因系列及Wnt抑制因子(WIF1)等表达变化。
结果与结论：与阴性对照组比较，诱导3 d后Fzd2表达升高11.86倍，Fzd3升高达到2倍；诱导7 d后，Fzd2表达升高5.12倍，Fzd3恢复到正常水平；β-catenin 在诱导3 d时表达升高达2倍；WIF1在诱导3 d和7 d后表达显著下降。结果提示Wnt信号途径可能参与了杜仲促骨髓间充质干细胞成骨分化过程。

关键词: 杜仲, 骨髓间充质干细胞, Wnt, 基因表达, 大鼠, 干细胞

Abstract:

BACKGROUND: Recently, there are many reports describing Chinese medicine and the effective components of Chinese medicine for treatment of osteoporosis. But there are few reports regarding cellular osteogenic differentiation and regulation signaling.
OBJECTIVE: To investigate the expression of Wnt signaling pathway related genes during osteogenic differentiation of rat bone marrow mesenchymal stem cells induced by eucommia bark.
METHODS: Passage 3 Sprague-Dawley rat bone marrow mesenchymal stem cells were inoculated into 6-well plate, with 1×103 cells per well. 24 hours later, induced medium supplemented with 7.5% fetal bovine serum-containing DMEM/F12 (1:1) and 1/1 000 eucommia bark was refreshed. Normal culture medium was used in the negative control group. After induction for 8 hours, 1, 3, and 7 days, expression levels of Fzd and LRP receptors of Wnt passway, β-catenin, intranuclear Wnt target genes and Wnt inhibitory factor were determined by RT-qPCR.
RESULTS AND CONCLUSION: Compared with negative control group, Fzd2 expression increased to 11.86 folds and Fzd3 expression increased to 2 folds respectively after induction for 3 days; Fzd2 expression increased to 5.12 folds and Fzd3 rebounded to normal level after induction for 7 days; β-catenin expression increased to 2 folds after induction for 3 days, and WIF1 expression decreased significantly after induction for 3 and 7 days, respectively. These findings suggest that Wnt signaling pathway may participate in the process of eucommia bark promoting in osteogenic differentiation of bone marrow mesenchymal stem cells.

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