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| 波尔定对破骨细胞分化及骨吸收功能的影响 |
| The effect of boldine on differentiation and resorption function of osteoclasts |
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| DOI：10.3969/j.issn.1006.7108.2016.03.004 |
| 中文关键词: 波尔定；破骨细胞 骨吸收 小鼠单核细胞 核因子kB受体活化因子 |
| 英文关键词:Boldine Osteoclast Bone resorption Mouse monocyte ceils Receptor activator of NF-kB Ligand |
| 基金项目:国家自然科学基金（81373773，81573845);北京市自然基金(7142144);中国中医科学院自主选题（YZ-1405) |
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| 中文摘要: |
| 目的 研究波尔定对NF-kB受体活化因子配体（RANKL)诱导的小鼠单核细胞（RAW264.7)向破骨细胞（OC)分化及骨吸收功能的影响。方法 采用RANKL(50 ng/ml)诱导RAW264.7细胞向OC分化，不同浓度波尔定（1、10、20、40、80μmol/ L)联合分化培养。培养第4d采用抗酒石酸酸性磷酸酶（TRAP)染色,计算OC样细胞数量；比色法测定TRAP活性；Real-time PCR法检测各组OC标志性基因核因子kB受体活化因子（RANK)、活化T细胞核因子l(NFATcl)和c-fos基因的表达;CCK-8 法检测波尔定对OC的毒性。培养至第9d,采用甲苯胺蓝染色并用Leica Qwin图像分析系统计算骨吸收陷窝面积。结果 与对照组比较,TRAP阳性OC数量、TRAP活性、OC标志性基因RANK、NFATcl和c-fos的表达量及骨陷窝面积，随着波尔定浓度增加而减少，与对照组比较，20μmol/ L、40μmol/ L 、80μmol/ L波尔定可明显减少OC数量和TRAP活性（P <0.05，P <0.01）,下调OC标志性基因RANK、NFATcl和c-fos的表达量及骨陷窝面积（P < 0.05),但以上浓度波尔定对OC均未产生毒 性。结论 波尔定对RANKL诱导的RAW264.7细胞向OC的分化及骨吸收功能具有抑制作用。 |
| 英文摘要: |
| Objective To investigate the effect of boldine on differentiation and resorption function of osteoclasts induced from mouse monocyte RAW264. 7 cells in the presence of receptor activator of NF-kB Ligand (RANKL). Methods The mouse monocyte RAW264. 7 cells were cultured in the presence of receptor activator of RANKL and with different concentration of boldine (1, 10, 20, 40, 80 μmol/L). On the 4 day, cells were stained by tartrate-resistant acid phosphatase (TRAP) and measured the TRAP activity to compare the number and activity of osteoclasts. CCK-8 method was used identify if boldine had toxicity on osteoclasts. The expression levels of receptor activator of NF-kB (RANK)，nuclear factor of activated T cells cl (NFATcl) and c?fos were detected by Real-time PCR. On the 9 day, the bone slices were stained by toluidine blue and quantified the resorption area by Leica Qwin system. Results Compared with control group, the number of osteoclasts was decreased with the increasing of boldine concentration. The groups of 20μmol/L, 40μmol/L and 80μmol/L boldine had significant differences (P < 0. 05, P < 0. 01), TRAP activity were also obviously decreased and showed no toxicity with the increasing concentration. Compared with control group, the resorption areas were significantly reduced when boldine concentration was increased. Meanwhile, The groups of 20μmol/L, 40μmol/L and 80μmol/L boldine had significant differences (P<0. 05). The expression levels of RANK,NFATcl and c-fos were down-regulated with the increasing of boldine concentration，and the groups of 20μmol/L, 40μmol/L and 80μmol/L boldine had significant differences ( P < 0.05). Conclusion Boldine could suppress the differentiation and resorption function of osteoclasts induced from RAW264.7 cells in the presence of RANKL. |
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