[1]赵志虎 李风波 马信龙 孙晓雷 马剑雄 张杨 王颖 卢斌 孙磊 王建宝 吕建伟.柚皮苷对共培养体系中成骨细胞活性及破骨细胞分化的影响[J].中华老年骨科与康复电子杂志,2015,(02):13-18.[doi:10.3877/cma.j.issn.2096-0263.2015.02.003]
　Zhao Zhihu,Li Fengbo,Ma Xinlong,et al.Effect of naringin on the activity of osteoblast and differentiation of osteoclast in a co-culture system[J].Chin J Geriatr Orthop Rehabil(Electronic Edition),2015,(02):13-18.[doi:10.3877/cma.j.issn.2096-0263.2015.02.003]
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柚皮苷对共培养体系中成骨细胞活性及破骨细胞分化的影响()
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中华老年骨科与康复电子杂志[ISSN:1674-3911/CN:11-9292/R]

卷:
期数:
2015年02期
页码:
13-18
栏目:
骨与创伤
出版日期:
2015-11-05

文章信息/Info

Title:
Effect of naringin on the activity of osteoblast and differentiation of osteoclast in a co-culture system
作者:
赵志虎 李风波 马信龙 孙晓雷 马剑雄 张杨 王颖 卢斌 孙磊 王建宝 吕建伟
天津医院骨科研究所生物力学研究室，骨科，骨科研究所形态学与材料研究室，骨科研究所 细胞与组织工程研究室，骨科研究所数字骨科研究室，骨科研究所影像学与体内示踪研究室
Author(s):
Zhao Zhihu;  Li Fengbo;  Ma Xinlong;  Sun Xiaolei;  Ma Jianxiong;  Zhang Yang;  Wang Ying;  Lu Bin;  Sun Lei;  Wang Jianbao;  Lyu Jianwei
Biomechanics Research of Institute of Orthopedics, Department of Orthopaedics, Morphology and Material Laboratory Research, Cell and Tissue Engineering Laboratory Research, Digital Orthopaedic Research, Imaging and Tracer of Laboratory, Tianjin Hospital
关键词:
柚皮苷; 　共同培养技术; 　成骨细胞; 　破骨细胞
Keywords:
Naringin;  　Coculture techniques;  　Osteoblasts;  　Osteoclasts
DOI:
10.3877/cma.j.issn.2096-0263.2015.02.003
摘要:
目的　研究在成骨细胞- 破骨细胞共培养体系中，柚皮苷对成骨细胞活性和破骨细 胞分化的影响。方法　将成骨细胞（MC3T3-E1 细胞株）和破骨细胞（RAW264.7 细胞株）以2 ∶ 1 的 比例分别培养至Transwell 小室的上室和下室。根据培养基是否含有柚皮苷分为对照组和柚皮苷（2 ng/ml 组、20 ng/ml 组、200 ng/ml 组），培养7 d 后对下室破骨细胞进行TRAP 染色和骨吸收陷窝鉴 定；荧光定量PCR 分析成骨细胞骨保护素（osteoprotegerin，OPG）、核因子κB 受体活化因子配体 （receptor activator for nuclear factor-κB ligand，RANKL）基因和破骨细胞分化基因活化T 细胞核因子-1 （NFATc-1）、活化T 细胞核因子-2（NFATc-2）和核因子κB 受体激活蛋白（receptor activator of NF- κB，RANK）的相对表达量。结果　与对照组相比，柚皮苷可以促进成骨细胞OPG、RANKL 的表 达量且可提高OPG/RANKL 的比值，差异有统计学意义（P ＜ 0.05）；20 ng/ml 柚皮苷TRAP（+） 细胞数（5.82±3.37）明显少于对照组（20.56±7.69），差异有统计学意义（P ＜ 0.05）；柚皮苷可 抑制破骨细胞NFATc-1、NFATc-2 的表达，促进RANK 的表达，差异有统计学意义（P ＜ 0.05）。结论 柚皮苷可促进共培养条件体系中成骨细胞OPG 和RANKL 的表达以及抑制破骨细胞的分化，与上调 OPG/RANKL 的比值有关。
Abstract:
　Objective　To explore the effect of naringin on the osteoblast activity and differentiation of osteoclast in a co-culture system. Methods　Osteoblasts (MC3T3-E1 cell line) and osteoclasts (RAW264.7 cell line) were separately cultured in the upper chamber and lower chamber of Transwell with proportion of 2:1, and divided into control group and naringin group according to whether the culture medium containing naringin. Three different doses of naringin (2 ng/ml, 20 ng/ ml, 200 ng/ml) were applied in naringin group. After 7 days of culturing, the osteoclasts were processed for TRAP staining and bone resorption lacunae identification. Fluorescent quantitative PCR were used to semiquantitative analysize the relative expression level of OPG, RANKL, NFATc-1, NFATc-2 and RANK (receptor activator of NF-κB). Results　Compared to the control group, Naringin can promote the expression of OPG and RANKL and increase the ratio of OPG/RANKL in osteoblasts, with significant differences (P<0.05). The number of TRAP(+) cells in Naringin group (20 ng/ml) (5.82±3.37) was less than that of control group (20.56±7.69), and the difference was statistically significant (P<0.05). Naringin can inhibit the expression of NFATc-1 and NFATc-2 in osteoclasts, while promoting the expression of RANK, and the difference is statistically significant (P<0.05). Conclusion　Naringin can promote the expression OPG and RANKL of osteoblasts and inhibiting osteoclast differentiation in co-culture system, which may be related to the increasement of OPG/RANKL ratio.

参考文献/References:

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备注/Memo

备注/Memo:
天津中医药管理局重点资助项目（13123）；国家自然科学基金青年基金（81401792）；国家自然科学 基金青年基金（81501061）
更新日期/Last Update: 2015-12-30