中国组织工程研究

中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (6): 835-840.doi: 10.3969/j.issn.2095-4344.2014.06.003

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

α-玉米赤霉醇影响小鼠骨髓间充质干细胞的成骨分化

邹 斌1，宗少晖2，曾高峰3，方 晔4，高太行4   

  1. 广西医科大学，1七年制临床医学2007级研究生，3公共卫生学院，4研究生学院，广西壮族自治区南宁市 530021；2广西医科大学第一附属医院脊柱骨病外科，广西壮族自治区南宁市 530021
  • 修回日期:2013-12-14 出版日期:2014-02-05 发布日期:2014-02-05
  • 通讯作者: 宗少晖，教授，广西医科大学第一附属医院脊柱骨病外科，广西壮族自治区南宁市 530021
  • 作者简介:邹斌，男，1989年生，江西省抚州市人，汉族，广西医科大学在读硕士，主要从事脊柱骨病外科疾病的基础与临床研究。

Effects of alpha-zearalanol on the osteogenic differentiation of mouse bone marrow mesenchymal stem cells

Zou Bin1, Zong Shao-hui2, Zeng Gao-feng3, Fang Ye4, Gao Tai-hang4   

  1. 1Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China; 2Department of Spinal Surgery, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China; 3School of Public Health, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China; 4Graduate School of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Revised:2013-12-14 Online:2014-02-05 Published:2014-02-05
  • Contact: Zong Shao-hui, Professor, Department of Spinal Surgery, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • About author:Zou Bin, Studying for master’s degree, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China

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摘要：

背景：骨髓间充质干细胞具有向成骨细胞分化的能力，植物雌激素α-玉米赤霉醇对小鼠骨髓间充质干细胞的成骨分化可能有促进作用。

目的：探讨α-玉米赤霉醇对小鼠骨髓间充质干细胞向成骨细胞分化的影响。
方法：采用全骨髓培养差速贴壁法体外分离培养小鼠骨髓间充质干细胞，取第3代细胞分为6组：非诱导组加含体积分数为10%胎牛血清、1%双抗的IMDM普通培养基；空白诱导组仅加入等量成骨条件培养基(含   0.1 μmol/L 地塞米松、10 mmol/L β-甘油磷酸钠、50 μmol/L 维生素C)；阳性对照诱导组加入等量成骨条件培养基及10-8 mol/L雌二醇；高、中、低α-玉米赤霉醇组分别加入成骨条件培养基及10-5，10-6，10-7 mol/L梯度浓度的α-玉米赤霉醇。倒置显微镜下观察细胞形态变化，MTT实验测定细胞增殖状态绘制细胞生长曲线，酶联免疫吸附法测定碱性磷酸酶、骨钙素的表达。

结果与结论：非诱导组细胞呈长梭形，生长密集时有接触抑制，各诱导组的细胞增殖受促进，诱导后细胞呈三角形、多角形、不规则形状，细胞可重叠生长；非诱导组低表达碱性磷酸酶、骨钙素，低浓度(10-7 mol/L) α-玉米赤霉醇组和阳性对照诱导组高表达碱性磷酸酶、骨钙素，与空白诱导组相比，差异有非常显著性意义(P < 0.01)。结果说明α-玉米赤霉醇可以促进骨髓间充质干细胞的增殖，低浓度α-玉米赤霉醇可显著促进体外培养的小鼠骨髓间充质干细胞成骨分化。而上调碱性磷酸酶、骨钙素的表达量可能是α-玉米赤霉醇诱导促进骨髓间充质干细胞骨向分化的作用机制之一。


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, 骨髓间充质干细胞, α-玉米赤霉醇, 成骨细胞分化, 碱性磷酸酶, 骨钙素

Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells can differentiate into osteoblasts, and α-zearalanol can promote the osteogenic differentiation of mouse bone marrow mesenchymal stem cells.

OBJECTIVE: To observe the effects of α-zearalanol on the osteogenic differentiation of mouse bone marrow mesenchymal stem cells.
METHODS: Mouse bone marrow mesenchymal stem cells were isolated through whole bone marrow adherence method. Passage 3 cells were divided into non-induced group (cultured by the Iscove’s modified Dulbecco’s medium containing 10% fetal bovine serum and 1% penicillin and streptomycin), blank induced group (induced by the classic osteoblast-induced system with 0.1 μmol/L dexamethasone, 10 mmol/L β-glycerine sodium, and    50 μmol/L vitamin C), positive control induced group (induced by the classic osteoblast-induced system and   10-8 mol/L oestrogen), and low-, median-, high-concentration α-zearalanol groups (induced by the classic osteoblast-induced system and 10-7, 10-6, 10-5 mol/L α-zearalanol). The morphological changes of induced cells were observed by phase contrast microscopy, and the growth curves of cultured cells were plotted based on MTT results. The expression of alkaline phosphatase and osteocalcin were detected by ELISA.

RESULTS AND CONCLUSION: In the non-induced group, cells showed long fusiform shape mostly, and contact inhibition was observed while the cells were close-packed. After induced, cells proliferated effectively, showed triangle, astero-form, polygon or irregular shape, and clumps and multilayer of cells became evident. Compared to both the blank induced group, the alkaline phosphatase activity and osteocalcin expression were significantly lower in the non-induced group, but higher in the positive control induced group and low concentration α-zearalanol group (P < 0.01). These findings indicate that α-zearalanol can promote the proliferation of bone marrow mesenchymal stem cells, and enhance the expression of alkaline phosphatase and osteocalcin expression, especially in the low concentration group. This demonstrates that low concentration α-zearalanol obviously enhances the osteogenic differentiation of mouse bone marrow mesenchymal stem cells in vitro. Up-regulating the release of alkaline phosphatase and osteocalcin may be one of the mechanisms by which α-zearalanol promotes osteogenic differentiation of bone marrow mesenchymal stem cells.


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:  bone marrow, mesenchymal stem cells, phytoestrogens, cell differentiation, alkaline phosphatase, osteocalcin

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