雌激素对大鼠骨髓间充质干细胞增殖及成骨分化的影响

上海口腔医学 ›› 2014, Vol. 23 ›› Issue (6): 654-660.

雌激素对大鼠骨髓间充质干细胞增殖及成骨分化的影响

王洁, 张鹏, 代庆刚, 欧阳宁鹃, 江凌勇^*, 房兵^*

上海交通大学医学院附属第九人民医院·口腔医学院口腔颅颌面科,上海市口腔医学重点实验室,上海 200011

收稿日期:2014-01-24 修回日期:2014-03-18 出版日期:2014-12-20 发布日期:2015-01-08
通讯作者: 房兵,Tel: 021-23271699-5735,E-mail: braces_dr@hotmail.com; E-mail:jadewong19881113@hotmail.com
作者简介:王洁（1988-）,女,硕士研究生
基金资助:
国家自然科学基金（81371121,11342005,30901698,10972142）; 上海市自然科学基金（13ZR1423700 ); 上海交通大学医工交叉基金（YG2012MS40 ); 上海市科学技术委员会基础研究重点项目（12JC1405700 )

The effect of estrogen on proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells

WANG Jie, ZHANG Peng, DAI Qing-gang, OUYANG Ning-juan, JIANG Ling-yong, FANG Bing

Department of Oral and Craniomaxillofacial Science, Ninth People’s Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai Key Lab of Stomatology. Shanghai 200011, China

Received:2014-01-24 Revised:2014-03-18 Online:2014-12-20 Published:2015-01-08
Supported by:
National Natural Science Foundation of China (81371121, 11342005, 30901698, 10972142), Natural Science Foundation of Shanghai Municipality (13ZR1423700), Collaborative Foundation of Medical and Engineering Science of Shanghai Jiao Tong University (YG2012MS40), Key Basic Research Foundation of Science and Technology Committee of Shanghai Municipality (12JC1405700), "Chen Xing" Project from Shanghai Jiao Tong University and Innovative Research Team of Shanghai Municipal Education Commission

摘要/Abstract

摘要： 目的：应用17 β-雌二醇（E₂）作用于大鼠骨髓间充质干细胞,研究不同浓度雌激素对细胞增殖、凋亡及成骨分化的影响。方法：原代培养大鼠骨髓间充质干细胞（rat bone mesenchymal stem cells,rBMSCs）,加入0、10^-9和10^-7 mol/L不同浓度E₂,CCK-8及Annexin V/PI双染法检测E₂对细胞增殖与凋亡的影响。分别于成骨诱导后第1、3、5、7、11、14和21天,采用碱性磷酸酶（alkaline phosphatase,ALP）活性测定、ALP和钙结节染色,以及实时定量PCR等方法检测不同浓度E₂对细胞成骨向分化能力的影响。采用SPSS19.0软件包对数据进行统计学分析。结果：E₂对大鼠骨髓间充质干细胞增殖与凋亡未见明显影响,并能显著提高干细胞的成骨分化能力,浓度为10^-9 mol/L时促进作用最为明显。与对照组相比,加药组细胞ALP活性升高,钙结节形成增多,成骨相关基因（RUNX2、ALP、COL I、OCN）表达显著升高。结论：17 β-雌二醇能促进大鼠骨髓间充质干细胞成骨分化,体外浓度为10^-9 mol/L时,作用最为显著。

关键词: 雌激素, 骨髓间充质干细胞, 成骨分化

Abstract: PURPOSE: Different concentrations of 17 β-estradiol (E₂) were applied in the osteogenic differentiation of rat bone mesenchymal stem cells (BMSCs), and the proliferation and apoptosis of BMSCs were explored. METHODS: BMSCs were obtained from the femurs and tibias of SD rats. The proliferation curve was conducted to rBMSCs in culture medium containing 0, 10^-9, and 10^-7mol/L 17 β-estradiol by CCK-8 for 7 days. Annexin V and PI for flow cytometry were applied to detect the impact of E₂ on apoptosis of rBMSCs. After 1, 7, 11 and 14 days of osteogenic induction, the activity of alkaline phosphatase (ALP) was assayed; ALP staining was performed on day 7 and day 14; Alizarin red staining for calcium deposits was carried out on day 21. Concentrations of 0, 10^-9, and 10^-7mol/L 17 β-estradiol were administrated to rBMSCs for real-time PCR of osteogenic related genes on day 1, 3, 5, 7, 14, and day 21. The data was statistically analyzed using SPSS 19.0 software package. RESULTS: The effect of 17 β-estradiol on proliferation and apoptosis of rBMSCs was not obvious. However, after osteogenic induction, the ALP activity and Alizarin red staining were significantly stronger in the groups containing 17 β-estradiol. Especially, the use of 17 β-estradiol with the concentration of 10^-9 mol/L enhanced the expression of osteogenic related genes like RUNX2, ALP, COL I, and OCN, which was significantly higher than other groups. CONCLUSIONS: 17 β-estradiol promotes osteogenic differentiation of BMSCs in a dose-dependent pattern in vitro.

Key words: Estrogen, Bone mesenchymal stem cells, Osteogenic differentiation

中图分类号:

R739.8

王洁, 张鹏, 代庆刚, 欧阳宁鹃, 江凌勇, 房兵. 雌激素对大鼠骨髓间充质干细胞增殖及成骨分化的影响[J]. 上海口腔医学, 2014, 23(6): 654-660.

WANG Jie, ZHANG Peng, DAI Qing-gang, OUYANG Ning-juan, JIANG Ling-yong, FANG Bing. The effect of estrogen on proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells[J]. Shanghai Journal of Stomatology, 2014, 23(6): 654-660.

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