Osteoprotective effects of osthole in a mouse model of 5/6 nephrectomy through inhibiting osteoclast formation
Osteoprotective effects of osthole in a mouse model of 5/6 nephrectomy through inhibiting osteoclast formation
复制标题
蛇床子素通过抑制破骨细胞形成对 5/6 肾切除小鼠模型的骨保护作用
DOI：
10.3892/mmr.2016.5687
复制
发表时间：
2016-10-01
影响因子：
3.4
通讯作者：
Shu, Bing
中科院分区：
文献类型：
--
作者：
Li, Xiaofeng;Xue, Chunchun;Shu, Bing
本研究旨在研究蛇床子素对5/6肾切除小鼠模型破骨细胞形成和骨丢失的影响。对照组和蛇床子素组的小鼠在5/6肾切除术后1个月分别用安慰剂或蛇床子素治疗。在肾切除术后2个月，收获L4椎骨。用显微CT测量松质骨的骨密度（BMD），用抗酒石酸酸性磷酸酶（TRAP）染色评价破骨细胞的形成。采用免疫组化染色和逆转录-定量聚合酶链反应（RT-PCR）检测活化T细胞核因子-胞浆蛋白-1（NFATc-1）、c-Fos、组织蛋白酶K、Trap、基质金属蛋白酶9（Mmp 9）、骨保护素（Opg）和核因子B受体激活因子配体（Rankl）的表达。骨髓细胞与蛇床子素共同培养，TRAP染色显示破骨细胞形成。原代培养的颅骨成骨细胞加入蛇床子素，检测成骨细胞中Opg和Rankl的表达水平。与假手术组比较，模型组小鼠骨密度明显降低。破骨细胞数量及NFATc-1、c-Fos、组织蛋白酶K和Mmp 9的表达水平显著增加。与对照组相比，蛇床子素组小鼠L4椎骨BMD增加，破骨细胞数量减少，NFATc-1，c-Fos，组织蛋白酶K和Mmp 9的表达水平降低。体外实验也表明，蛇床子素处理后，破骨细胞的形成减少。骨保护素（Opg）/核因子-B配体（Rankl）的受体激活剂上调蛇床子素治疗L4椎骨和原代培养的颅骨成骨细胞。蛇床子素通过上调OPG/RANKL抑制破骨细胞的形成，部分逆转5/6肾切除小鼠骨丢失。
The present study aimed to investigate the effects of osthole on osteoclast formation and bone loss in a mouse model of 5/6 nephrectomy. The mice in control and osthole groups were treated 1 month following 5/6 nephrectomy with either a placebo or osthole, respectively. At 2 months post-nephrectomy, the L4 vertebrae were harvested. The bone mineral density (BMD) of cancellous bone was measured using micro-CT and tartrate-resistant acid phosphatase (TRAP) staining was performed to evaluate osteoclast formation. Immunohistochemistry staining and reverse transcription-quantitative polymerase chain reaction were performed to detect the expression of nuclear factor of activated T-cells, cytoplasmic-1 (NFATc-1), c-Fos, cathepsin K, Trap, matrix metalloproteinase 9 (Mmp9), osteoprotegerin (Opg) and receptor activator for nuclear factor-B ligand (Rankl). Bone marrow cells were cultured with osthole, and osteoclast formation was shown by TRAP staining. Primary calvaria osteoblasts were cultured with osthole, and expression levels of Opg and Rankl were detected. Compared with the sham group, the BMD of mice in model group was significantly reduced. The numbers of osteoclasts and the expression levels of NFATc-1, c-Fos, cathepsin K and Mmp9 were significantly increased. Compared with the control group, the mice in the osthole group exhibited increased BMD of the L4 vertebrae, a reduction in osteoclast numbers and decreased expression levels of NFATc-1, c-Fos, cathepsin K and Mmp9. In vitro experiments also showed that osteoclast formation was decreased following treatment with osthole. Osteoprotegerin (Opg)/receptor activator for nuclear factor-B ligand (Rankl) was upregulated by osthole treatment in the L4 vertebrae and in primary cultures of calvarial osteoblasts. Osthole inhibited osteoclast formation and partially reversed the bone loss induced by 5/6 nephrectomy in mice through the upregulation of OPG/RANKL.