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

作　　者：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] WANG Sihan JIANG Huan YU Hang LIU Shaowei LI Menghong WANG Liuyi YANG Junxing HU Min(Department of Orthodontics, Stomatology Hospital, Jilin University, Changchun 130021, China Jilin Provincial Kay Laboratory of Tooth Development and Bone Remodeling, Changchun 130021, China)

机构地区：[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

出　　处：《吉林大学学报（医学版）》2017年第2期236-240,I0001,共6页Journal of Jilin University：Medicine Edition

基　　金：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)

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5d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。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 RAW264.7were induced to the OC.Tacrolimus acid phosphatase（TRAP）staining and bone resorption lacunae observation were used to identify the induction.CCK-8method was used to select the appropriate concentration of UA for RAW264.7cell-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 40.0μmol·L^-1）were added.UA was not added in control group.Results：The TRAP staining and bone resorption lacunae observation showed that after the RAW264.7cells were induced for 5d,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 RAW264.7cells were successfully differentiated into the osteoclasts.The CCK-8detection results showed that high concentration of UA（ 10.0μmol·L^-1）significantly inhibited the proliferation of OC;the appropriate concentration of UA（5.0μmol·L^-1）was in the biological safety concentration range and could inhibit the OC proliferation;low concentration of UA（2.5μmol·L^-1）had no effect.Conclusion：RANKL can induce the differentiation and maturation of RAW264.7cells.UA is correlated with the proliferation and differentiation of OC;UA has inhibitory effect on OC at the appropriate concentration（5.0μmol·L^-1）in a time-dependent manner.

关 键 词：熊果酸 破骨细胞 RAW264.7细胞 牙根吸收 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]