天津医药

天津医药 ›› 2018, Vol. 46 ›› Issue (2): 122-125.doi: 10.11958/20171264

• 细胞与分子生物学 • 上一篇    下一篇

不同浓度西红花苷对RANKL诱导的小鼠破骨细胞分化的影响

林思思1 ， 章礼炜2 ， 孙旺1 ， 施更生1△   

  1. 1临海， 温州医科大学附属浙江省台州医院口腔科 （邮编317000）， 2骨科
  • 收稿日期:2017-11-14 修回日期:2017-12-20 出版日期:2018-02-15 发布日期:2018-02-15
  • 通讯作者: 施更生 E-mail:shigs@enzemed.com

The effects of different concentrations of crocin on receptor activator of nuclear factor kappa B ligand (RANKL)–induced osteoclastogenesis

LIN Si-si 1 , ZHANG Li-wei 2 , SUN Wang1 , SHI Geng-sheng1△   

  • Received:2017-11-14 Revised:2017-12-20 Published:2018-02-15 Online:2018-02-15

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摘要： 摘要： 目的 研究不同浓度的西红花苷对核因子 κB 受体活化因子配体 （RANKL） 诱导的破骨细胞分化的影响。方法 常规培养小鼠巨噬细胞 RAW264.7， 经 0、 6.25、 12.5、 25、 50、 100、 200、 400 μmol/L 浓度的西红花苷处理后， 采用 CCK-8 法检测细胞生长活力的变化， 筛选出合适的实验浓度。RAW264.7 细胞经 RANKL （100 ng/L） 诱导形成破骨细胞系后， 分别经 0、 12.5、 50、 100 μmol/L 的西红花苷处理， 抗酒石酸酸性磷酸酶 （TRAP） 染色观察各浓度组破骨细胞的生成数量； 实时定量 PCR （real-time PCR） 检测破骨细胞的特异性基因降钙素受体 （CTR）、 活化 T 细胞核因子 1 （NFATC1）、 C-fos、 TRAP mRNA 的表达水平变化。结果 在 0~100 μmol/L 范围内， 西红花苷对 RAW264.7 细胞的生长无明显影响 （P＞0.05）。当西红花苷浓度≥100 μmol/L 时， 细胞增殖活力下降， 显示出明显的抑制增殖作用 （P＜ 0.05）， 选取 0~100 μmol/L 西红花苷进行实验。随着西红花苷的处理浓度的升高， RAW264.7 分化成破骨细胞的数量明显减少， 同时破骨细胞的特异性基因 CTR、 NFATC1、 C-fos 和 TRAP mRNA 的表达水平也明显下降 （P＜0.05）。结论 在一定浓度范围内 （0~100 μmol/L）， 高水平的西红花苷可明显抑制破骨细胞的分化。

关键词: 关键词： 西红花△, 巨噬细胞, 细胞分化, 破骨细胞, 牙周炎, 细胞增殖, 西红花苷, 核因子 κB 受体活化因子配体

Abstract: Abstract: Objective To investigate the effects of different concentrations of crocin on receptor activator of nuclear factor kappa B ligand (RANKL) - induced osteoclastogenesis using the monocyte-macrophage cell line RAW264.7. Methods The monocyte-macrophage cell line RAW264.7 was cultured routinely. After treatment with 0, 6.25, 12.5, 25, 50, 100, 200 and 400 μmol/L crocin, cell counting kit-8 (CCK-8) assay was used to analyze the viability of RAW264.7 cells to screen out the appropriate experimental concentration. RAW264.7 cells were induced by RANKL (100 ng / L) to form osteoclasts. After treated with 0, 12.5, 50 and 100 μmol / L crocin respectively, the number of osteolasts was counted by tatrate resistant acid phosphatasec (TRAP) staining. Real-time PCR was used to detect the mRNA expression levels of calcitonin receptor (CTR), nuclear factor of active T cells 1 (NFATC1), C-fos and TRAP. Results No significant effects of crocin (within 0-100 μmol / L) were found on the viability of RAW264.7 cells (P＞0.05). However, When crocin concentration was over 100 μmol/L, the cell proliferation was decreased, and which showed a significant inhibitory effect on proliferation (P＜0.05). Thus, 0-100 μmol / L crocin was selected as the experiment concentration. The amount of differentiated osteolasts and the expression levels of CTR, NFATC1, C-fos and TRAP mRNA were decreased significantly with the increased concentrations of crocin (P＜0.05). Conclusion At a certain concentration (0-100 μmol/L), the higher levels of crocin could inhibit RANKL-induced osteoclastogenesis.

Key words: Key words：CROCUS SATIVUS, macrophages, cell differentiation, osteoclasts, periodontitis, cell proliferation, crocin, receptor activator of nuclear factor kappa B ligand