| 蛇床子素对体外培养成骨细胞增殖与分化成熟的影响 |
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投稿时间：2010-03-23
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| 作者 | Author | 单位 | Address | E-Mail |
| 明磊国 |
MING Lei-guo |
兰州军区总医院骨研所，甘肃 兰州 730050 |
Institute of Orthopaedics,Lanzhou General Hospital of PLA,Lanzhou 730050,Gansu,China |
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| 葛宝丰 |
GE Bao-feng |
兰州军区总医院骨研所，甘肃 兰州 730050 |
Institute of Orthopaedics,Lanzhou General Hospital of PLA,Lanzhou 730050,Gansu,China |
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| 陈克明 |
CHEN Ke-ming |
兰州军区总医院骨研所，甘肃 兰州 730050 |
Institute of Orthopaedics,Lanzhou General Hospital of PLA,Lanzhou 730050,Gansu,China |
chkeming@yahoo.com.cn |
| 马慧萍 |
MA Hui-ping |
兰州军区总医院药材科 |
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| 翟远坤 |
ZHAI Yuan-kun |
兰州军区总医院骨研所，甘肃 兰州 730050 |
Institute of Orthopaedics,Lanzhou General Hospital of PLA,Lanzhou 730050,Gansu,China |
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| 周建 |
ZHOU Jian |
兰州军区总医院骨研所，甘肃 兰州 730050 |
Institute of Orthopaedics,Lanzhou General Hospital of PLA,Lanzhou 730050,Gansu,China |
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| 李志锋 |
LI Zhi-feng |
兰州军区总医院骨研所，甘肃 兰州 730050 |
Institute of Orthopaedics,Lanzhou General Hospital of PLA,Lanzhou 730050,Gansu,China |
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| 期刊信息：《中国骨伤》2010年，第23卷，第9期，第688-691页 |
| DOI：10.3969/j.issn.1003-0034.2010.09.015 |
| 基金项目:甘肃省科技计划资助项目(编号：092nkDA025) |
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中文摘要:
目的：研究蛇床子素对体外培养大鼠成骨细胞(rat calvarial osteoblasts,ROB)增殖与分化成熟的影响。
方法：取新生SD大鼠颅骨，多次酶消化法获得成骨细胞，培养于含10%FBS的MEM培养液中，3 d后首次换液，铺满90%皿底后传代培养。增殖分析采用96孔板MTT法分析，加入培养液中终浓度分别为1×10-4、1×10-5、1×10-6、1×10-7 mol/L的蛇床子素；分化分析采用24孔板，于成骨性诱导培养第3、6、9、12、15天分别测碱性磷酸酶(alkaline phosphatase,ALP)活性，第8天用免疫组织化学法观察Ⅰ型胶原合成情况，第12天进行ALP组织化学染色，第14天进行茜素红染色和钙化结节计数。
结果：终浓度为1×10-4 mol/L蛇床子素对细胞增殖有抑制作用，而1×10-5 mol/L浓度虽对ROB增殖无明显影响，但能显着提高ALP活性，促进Ⅰ型胶原表达，并增加钙化结节数量。
结论：1×10-5 mol/L蛇床子素能促进ROB分化成熟，应为中药蛇床子抗骨质疏松的有效成分。 |
| 【关键词】成骨细胞 细胞培养技术 蛇床子素 骨质疏松 |
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| Effect of Osthol on the proliferation and differentiate of Osteoblasts in vitro |
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ABSTRACT
Objective: To investigate the effects of Osthol on the proliferation and differentiation of osteoblasts of rats(rat calvarial osteoblasts,ROB) cultured in vitro.
Methods: The neonatal SD rat skull was segregated,and enzyme digestion was used to obtain bone cells which were cultured in MEM containing 10% FBS. The medium was changed every three days,and serial subcultivation was performed when cells covered with 90% of the culture dish. The Osthol was added to 96-well plates with final concentration of 1×10-4 mol/L,1×10-5 mol/L,1×10-6 mol/L and 1×10-7 mol/L,and MTT method was used to evaluate the proliferation. Differentiation analysis:the alkaline phosphatase (ALP) activity was determined at the 3rd,6th,9th,12th and 15th days separately after osteogenic induction culture. The synthesis of typeⅠcollagen was observed using immunohistochemical method at the 8th day. The ALP stain was performed at the 12th day. The alizarin red staining was done and calcified nodules was counted at the 14th day.
Results: The Osthol with final concentration of 1×10-4 mo/L inhibit the proliferation of ROB. The Osthol with final concentration of 1×10-5 mol/L had no obvious influence on the proliferation of ROB,but it significantly promoted the activity of ALP,enhanced the synthesis of collagen type Ⅰ and increased the number of calcified nodules.
Conclusion: The Osthol with final concentration of 1×10-5 mol/L can promote differentiation and maturation of ROB,which may be active ingredients of Chinese drugs for the osteoporosis prophylaxis. |
| KEY WORDS Osteoblasts Cell culture techniques Osthole Osteoporosis |
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| 中文格式: | 明磊国,葛宝丰,陈克明,马慧萍,翟远坤,周建,李志锋.蛇床子素对体外培养成骨细胞增殖与分化成熟的影响[J].中国骨伤,2010,23(9):688~691 |
| 英文格式: | MING Lei-guo,GE Bao-feng,CHEN Ke-ming,MA Hui-ping,ZHAI Yuan-kun,ZHOU Jian,LI Zhi-feng.Effect of Osthol on the proliferation and differentiate of Osteoblasts in vitro[J].zhongguo gu shang / China J Orthop Trauma ,2010,23(9):688~691 |
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