不同浓度熊果酸对破骨细胞增殖分化的作用及其意义

doi:10.13481/j.1671-587x.20170205

吉林大学学报(医学版) ›› 2017, Vol. 43 ›› Issue (02): 236-240.doi: 10.13481/j.1671-587x.20170205

不同浓度熊果酸对破骨细胞增殖分化的作用及其意义

王思涵^1,2, 姜欢¹, 于航¹, 刘少伟¹, 李梦红¹, 王六一¹, 杨军星¹, 胡敏¹

1. 吉林大学口腔医院正畸科, 吉林长春 130021;
2. 吉林省牙发育及颌骨重塑与再生重点实验室, 吉林长春 130021

收稿日期:2016-12-21 出版日期:2017-03-28 发布日期:2017-03-31
通讯作者: 胡敏,教授,博士研究生导师(Tel:0431-88796023,E-mail:humin@jlu.edu.cn) E-mail:humin@jlu.edu.cn
作者简介:王思涵(1989-),女,黑龙江省牡丹江市人,在读医学硕士,主要从事正畸与牙根吸收方面的研究。
基金资助:
国家自然科学基金资助课题（81600898）；吉林省卫生厅科研基金资助课题（2014Z073）

Effects of ursolic acid at different concentrations on proliferation and differentiation of osteoclasts and their significances

WANG Sihan^1,2, JIANG Huan¹, YU Hang¹, LIU Shaowei¹, LI Menghong¹, WANG Liuyi¹, YANG Junxing¹, HU Min¹

1. Department of Orthodontics, StomatologyHospital, Jilin University, Changchun 130021, China;
2. Jilin Provincial Kay Laboratory of Tooth Development and Bone Remodeling, Changchun 130021, China

Received:2016-12-21 Online:2017-03-28 Published:2017-03-31

摘要/Abstract

摘要： 目的：研究熊果酸（UA）对破骨细胞（OC）增殖分化的影响，探讨其在正畸力致牙根吸收过程中的作用以及与OC间的关系。方法：对小鼠单核/巨噬系细胞RAW264.7进行OC诱导，采用抗酒石酸酸性磷酸酶（TRAP）染色法及骨吸收陷窝观察鉴定诱导是否成功。采用CCK-8法选取对RAW264.7无细胞生物毒性适宜浓度的UA，观察其对RAW264.7增殖分化的影响。实验组在培养液中加入梯度浓度为1.0、2.5、5.0、10.0、20.0和40.0μmol·L^-1 UA，对照组不加UA。结果：TRAP染色及骨吸收陷窝观察，诱导培养RAW264.7细胞5 d后，可见TRAP染色阳性细胞；扫描电镜下吸收陷窝呈圆形和椭圆形等，底壁较粗糙，边界较清晰。表明RAW264.7细胞成功向OC分化。CCK-8检测，高浓度UA（>10.0μmol·L^-1）显著抑制OC的增殖；适宜浓度UA（5.0μmol·L^-1）既在生物安全浓度范围内，又抑制OC的增殖；低浓度UA（< 2.5μmol·L^-1）无作用。结论：RANKL可诱导RAW264.7细胞分化成熟。UA与OC增殖分化有关联，其在适宜浓度（5.0μmol·L^-1）时对OC增殖有抑制作用，且呈时间依赖性。

关键词: 熊果酸, 牙根吸收, RAW264.7细胞, 破骨细胞

Abstract: Objective: To investigate the effects of ursolic acid (UA) on the proliferation and differentiation of osteoclasts (OC), and to explore its role in orthodontic force-induced root resorption and its relationship with OC. Methods: The mononuclear/macrophage cells RAW264.7 were induced to the OC.Tacrolimus acid phosphatase (TRAP) staining and bone resorption lacunae observation were used to identify the induction. CCK-8 method was used to select the appropriate concentration of UA for RAW264.7 cell-free biotoxicity and to observe its effect on the proliferation and differentiation of RAW264.7.In experimental groups, UA with gradient concentrations (1.0,2.5,5.0,10.0,20.0and 40.0 μmol·L^-1)were added.UA was not added in control group. Results: The TRAP staining and bone resorption lacunae observation showed that after the RAW264.7 cells were induced for 5 d, the TRAP staining positive cells were found; the resorption lacunae were rounded,and oval, etc,the bottom wall was coarser,and the boundary was clear,which indicated that the RAW264.7 cells were successfully differentiated into the osteoclasts.The CCK-8 detection results showed that high concentration of UA (> 10.0 μmol·L^-1) significantly inhibited the proliferation of OC;the appropriate concentration of UA (5.0 μmol·L^-1) was in the biological safety concentration range and could inhibit the OC proliferation; low concentration of UA (<2.5 μmol·L^-1) had no effect. Conclusion: RANKL can induce the differentiation and maturation of RAW264.7 cells. UA is correlated with the proliferation and differentiation of OC; UA has inhibitory effect on OC at the appropriate concentration (5.0 μmol·L^-1) in a time-dependent manner.

Key words: osteoclasts, ursolic acid, root resorption, RAW264.7 cells

中图分类号:

R329.2

王思涵, 姜欢, 于航, 刘少伟, 李梦红, 王六一, 杨军星, 胡敏. 不同浓度熊果酸对破骨细胞增殖分化的作用及其意义[J]. 吉林大学学报(医学版), 2017, 43(02): 236-240.

WANG Sihan, JIANG Huan, YU Hang, LIU Shaowei, LI Menghong, WANG Liuyi, YANG Junxing, HU Min. Effects of ursolic acid at different concentrations on proliferation and differentiation of osteoclasts and their significances[J]. Journal of Jilin University Medicine Edition, 2017, 43(02): 236-240.

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不同浓度熊果酸对破骨细胞增殖分化的作用及其意义

Effects of ursolic acid at different concentrations on proliferation and differentiation of osteoclasts and their significances

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