| 蛇床子素对新生大鼠成骨细胞增殖的影响及相关机制研究 |
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投稿时间：2012-08-31
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| 作者 | Author | 单位 | Address | E-Mail |
| 李玲慧 |
LI Ling-hui |
上海中医药大学附属曙光医院石氏伤科医学中心, 上海 201203
上海市中医药研究院骨伤科研究所, 上海 201203 |
Shi's Center of Orthopaedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai 201203, China |
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| 丁道芳 |
DING Dao-fang |
上海中医药大学附属曙光医院石氏伤科医学中心, 上海 201203
上海市中医药研究院骨伤科研究所, 上海 201203 |
Shi's Center of Orthopaedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai 201203, China |
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| 杜国庆 |
DU Guo-qing |
上海中医药大学附属曙光医院石氏伤科医学中心, 上海 201203
上海市中医药研究院骨伤科研究所, 上海 201203 |
Shi's Center of Orthopaedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai 201203, China |
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| 龚浩 |
GONG Hao |
中国中医科学院望京医院, 北京 100102 |
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| 王辉昊 |
WANG Hui-hao |
上海中医药大学附属曙光医院石氏伤科医学中心, 上海 201203
上海市中医药研究院骨伤科研究所, 上海 201203 |
Shi's Center of Orthopaedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai 201203, China |
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| 詹红生 |
ZHAN Hong-sheng |
上海中医药大学附属曙光医院石氏伤科医学中心, 上海 201203 |
Shi's Center of Orthopaedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai 201203, China |
zhanhongsheng2010@163.com |
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| 期刊信息：《中国骨伤》2013年，第26卷，第5期，第419-422页 |
| DOI：10.3969/j.issn.1003-0034.2013.05.017 |
| 基金项目:国家自然基金项目(编号:81073114,81001527,81102603,81173621);“中医骨伤科学”国家重点学科(编号:100508);上海领军人才项目(编号:041);上海高校“中医脊柱病损研究”创新团队建设项目(编号:2009-26);上海市科委重点项目(编号:09dZ1973800,08411950300,11DZ1973400,11XD1404600) |
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中文摘要:
目的: 探讨蛇床子素对体外培养的大鼠成骨细胞增殖的影响及机制。
方法: 新生SD大鼠10只, 脱颈处死后取出颅盖骨, 剪成约1 mm×1 mm的碎片, 采用胰酶预消化15 min,继以0.1 %Ⅰ型胶原酶37℃消化2次, 每次1 h,所得细胞混匀后置于5% CO2培养箱中37℃恒温培养。细胞培养48 h、28 d后行ALP染色和矿化结节染色以进行成骨性鉴定。细胞随机分为5组, 分别加入终浓度为100、50、25、12.5、0 μmol/L的蛇床子素进行干预。24、48、72 h后采用CCK-8法检测细胞增殖率, 并通过Western Blot方法检测加药1周后PCNA、β-catenin蛋白表达量。
结果: 镜下观察细胞形态不规则, 具有典型的成骨细胞形态;ALP染色及矿化结节染色均为阳性。CCK-8检测发现100 μmol/L的蛇床子素干预24 h后即可抑制成骨细胞生长, 50 μmol/L和25 μmol/L的蛇床子素于加药后48 h开始抑制成骨细胞生长, 12.5 μmol/L蛇床子素对成骨细胞的增殖无显着影响。Western Blot检测结果显示PCNA及β-catenin随蛇床子素浓度增加而表达下调。
结论: 100、50、25 μmol/L各个浓度的蛇床子素可抑制成骨细胞增殖, 12.5 μmol/L蛇床子素对细胞的增殖特性无显着影响。蛇床子素抑制成骨细胞增殖的作用可能是通过干扰Wnt/β-catenin信号通路而实现的。 |
| 【关键词】蛇床子 成骨细胞 大鼠 细胞增殖 |
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| Effect of osthole on proliferation of neonatal rat osteoblast and the relative mechanism research |
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ABSTRACT
Objective: To investigate the effects of osthole on proliferation of neonatal rat osteoblast and the mechanism.
Methods: Ten 24 hours old SD rats were executed by dislocating. The cranium of rats were removed and cut into blocks of 1 mm×1 mm size. After digested by trypsin for 15 min,the cranium were digested by type Ⅰcollagenase for one hour two times. The mixed cells were cultured in thermostat incubator with 5%CO2 under the condition of 37℃。 To identify the cells,ALP staining and alizarin red staining were performed after cultured 48 h and 28 d. The osteoblasts were randomly divided into five groups. Cells were treated with osthole at concentrations of 100,50,25,12.5,0 μmol/L. CCK-8 method was used to evaluate the proliferation after 24 h,48 h and 72 h. The expression of PCNA and β-catenin protein were detected through the method of Western Blot after one week.
Results: The cells had irregular shapes and showed typical features of osteoblast. The results of ALP staining and alizarin red staining were both positive. CCK-8 detection showed that the osthole with final concentration of 100 μmol/L inhibited the proliferation of osteoblast after 24 h,while the osthole with final concentrations of 50 μmol/L and 25 μmol/L displayed the inhibition effect after 48 h. The osthole of 12.5 μmol/L had no obvious influence on the proliferation of osteoblast. The result of Western Blot showed that osthole reduced the expression of PCNA and β-catenin protein in a dose-dependent manner.
Conclusion: The osthole with final concentrations of 100,50,25 μmol/L inhibited the proliferation of osteoblast (P<0.05). The osthole with final concentrations of 12.5 μmol/L had no obvious influence on the proliferation of osteoblast(P>0.05). These findings demonstrate that osthole may inhibit the proliferation of osteoblast by regulating the Wnt/β-catenin signaling in osteoblast. |
| KEY WORDS Fructus cnidii Osteoblasts Rats Cell proliferation |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 李玲慧,丁道芳,杜国庆,龚浩,王辉昊,詹红生.蛇床子素对新生大鼠成骨细胞增殖的影响及相关机制研究[J].中国骨伤,2013,26(5):419~422 |
| 英文格式: | LI Ling-hui,DING Dao-fang,DU Guo-qing,GONG Hao,WANG Hui-hao,ZHAN Hong-sheng.Effect of osthole on proliferation of neonatal rat osteoblast and the relative mechanism research[J].zhongguo gu shang / China J Orthop Trauma ,2013,26(5):419~422 |
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