天津医药

天津医药 ›› 2018, Vol. 46 ›› Issue (6): 624-628.doi: 10.11958/20180184

• 实验研究 • 上一篇    下一篇

灯心草抑制RANKL诱导的破骨细胞形成

王鹏，郭狄，陈利华，章礼炜，陈海啸   

  1. 温州医科大学附属台州医院骨科（邮编317000）
  • 收稿日期:2018-01-31 修回日期:2018-03-26 出版日期:2018-06-15 发布日期:2018-07-05
  • 通讯作者: 王鹏 E-mail:1181725936@qq.com

Juncus inhibits osteoclast formation induced by RANKL

WANG Peng, GUO Di, CHEN Li-hua, ZHANG Li-wei, CHEN Hai-xiao   

  1. Department of Orthopedics, Taizhou Hospital Affiliated to Wenzhou Medical University, Zhejiang 317000, China
  • Received:2018-01-31 Revised:2018-03-26 Published:2018-06-15 Online:2018-07-05

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摘要： 目的 观察灯心草对核因子（NF）-κB受体活化因子配体（RANKL）诱导骨髓巨噬细胞（BMMs）分化形成破骨细胞的作用，并探讨其机制。方法 取8周龄C57/BL6小鼠分离出BMMs，采用CCK-8法检测灯心草对BMMs增殖的影响。在30 μg/L巨噬细胞集落刺激因子（M-CSF）和50 μg/L RANKL刺激BMMs分化成破骨细胞，经不同浓度灯心草（0、6.25、12.5、25 μmol/L）处理，通过抗酒石酸酸性磷酸酶（TRAP）检测破骨细胞分化的情况。RT-PCR检测破骨细胞分化过程中特异性基因降钙素受体（CTR）、活化T细胞核因子1（NFATC1）、空泡型H+三磷酸腺苷转运酶-d2（VATPase-d2）、空泡型H+三磷酸腺苷转运酶-a3（V-ATPase-a3）、C-FOS的表达。结果 CCK-8实验结果显示，灯心草浓度低于12.5 μmol/L时对BMMs细胞增殖无明显抑制。50 μg/L RANKL可诱导BMMs细胞分化成TRAP染色阳性的多核巨细胞，即成熟的破骨细胞。不同浓度的灯心草可显著抑制破骨细胞形成，且呈现浓度依赖性；RT-PCR结果显示，灯心草浓度依赖性地抑制破骨细胞分化过程中特异性基因的表达。结论 灯心草浓度依赖性地抑制破骨细胞的形成，其机制可能是通过抑制破骨细胞分化过程中特异性基因的表达来实现的。

关键词: 灯心草, 破骨细胞, 骨质疏松, RANKL, TRAP

Abstract: Objective To observe the effect of Juncus effuses on osteoclasts differentiation from bone marrow macrophages (BMMs) induced by receptor activator for nuclear factor - κ B ligand (RANKL), and its mechanism thereof. Methods BMMs were isolated from whole bone marrow of 8-week-old C57/BL6 mice, and CCK-8 was used to detect the effect of Juncus on BMMs cell proliferation. Tartrate resistant acid phosphatase (TRAP) staining was used to show that 50 μg/L RANKL and 30 μg/L macrophage colony stimulating factor (M-CSF) stimulated the BMMs differentiation into osteoclasts, but the process was inhibited by Juncus (0, 6.25, 12.5 and 25 μmol / L). RT-PCR was used to detect the expressions of osteoclast-specific genes including calcitonin receptor (CTR), vacuolated H+ triphosphate transporter -d2 (V-ATPase-d2) and - a3 (V-ATPase-a3), activated T nuclear factor 1 (NFATC1) and C-FOS. Results There was no inhibition in the proliferation of BMMs cells treated with Juncus less than 12.5 mol/L detected by CCK-8. The 50 μg/L RANKL can induce BMMs differentiated into positive multinuclear giant cells detected by TRAP staining, but Juncus significantly inhibited osteoclast formation with a concentration dependence. The results of RT-PCR experiment showed that Juncus inhibited the expression of specific genes in osteoclast differentiation in concentration-dependent manner. Conclusion Juncus can inhibit osteoclast formation in concentration-dependent manner, resulting from the inhibitory effect on osteoclast specific gene expression.

Key words: Juncus effuses, osteoclasts, osteoporosis, RANKL, TRAP