在骨质疏松性骨折大鼠模型中，骨激酶下调基质Gla蛋白（Mgp）。

Osteoking downregulates Mgp in an osteoporotic fracture rat model.

作者信息

Sun Yan, Wang Xiaoqi, Chen Ran, Zhu Di, Shen Zhiqiang, Zhao Hongbin, Lee Wenhui

机构信息

Pharmaceutical College & Key Laboratory of Pharmacology for Natural Products of Yunnan Province, Kunming Medical University, Kunming Yunnan, 650500, China.

Key Laboratory of Bio-active Peptides of Yunnan Province/Key Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences, Kunming Institute of Zoology, Kunming Yunnan, 650032, China.

出版信息

J Tradit Chin Med. 2020 Jun;40(3):422-431. doi: 10.19852/j.cnki.jtcm.2020.03.010.

DOI:10.19852/j.cnki.jtcm.2020.03.010

PMID:32506856

Abstract

OBJECTIVE

To investigate the effectiveness of osteoking, a Traditional Chinese Medicine originating from Yi nationality, against osteoporosis (OP) and osteoporotic fracture (OPF), and to elucidate its mechanism of action.

METHODS

An osteoporotic fracture rat model was established; animals were divided into three treatment groups: parathyroid hormone, osteoking and 0.9%NaCl. After 4, 8 and 12 weeks of treatment, serum and bone tissues were collected. Enzyme-linked immuno sorbent assay, x-ray, histopathological evaluation and proteomics were used. Proteomics and GO annotation were performed based on identified peptides. The relative network was obtained from the STRING database and verified by polymerase chain reaction and Western blotting.

RESULTS

After osteoking treatment, the bone mineral density (BMD) increased with time in the osteoking group. At week 12, the BMD and bone mineral salt content of the osteoking group were 4.5% and 20.6% higher than those of the negative control group, respectively. Furthermore, the body weight followed the order of positive control group > osteoking group > negative control group, with significant differences among the groups (P < 0.05). Micro-CT analysis of femur sections revealed that the bone surface/volume ratio was significantly higher in the osteoking group than that in the negative control group. X-ray images demonstrated that the osteoking group showed clear callus. Moreover, high-voltage micro-CT demonstrated a massive cortical bone accumulation in the osteoking group. The gray values of callus in the osteoking group were higher than those in the negative group. From week 4 to 12, the serum bone alkaline phosphatase level increased by 49.6% in the osteoking group and the serum propeptide of type Ⅰprocollagen level decreased by 80.6%. Alizarin red staining demonstrated that the calcium deposition in the osteoking group was higher than that in the negative control group. Notably, the expression of Mgp, a key osteogenesis inhibitor, was lower in the osteoking group compared with the negative control group. Moreover, Sparc, bone morphogenetic protein-2 and Bglap expression was higher in the osteoking group through activation of the transforming growth factor-receptor activator of nuclear factor κB Ligand pathway.

CONCLUSION

Osteoking treatment increased bone quality and promoted calcium deposition. The results suggest that osteoking inhibits Mgp through the TGF-β/RANKL pathway to improve OP/OPF.

摘要

目的

研究源于彝族的中药骨刻灵对骨质疏松症（OP）和骨质疏松性骨折（OPF）的疗效，并阐明其作用机制。

方法

建立骨质疏松性骨折大鼠模型；将动物分为三个治疗组：甲状旁腺激素组、骨刻灵组和0.9%氯化钠组。治疗4、8和12周后，收集血清和骨组织。采用酶联免疫吸附测定、X射线、组织病理学评估和蛋白质组学方法。基于鉴定出的肽段进行蛋白质组学和基因本体注释。从STRING数据库获得相关网络，并通过聚合酶链反应和蛋白质免疫印迹法进行验证。

结果

骨刻灵治疗后，骨刻灵组骨密度（BMD）随时间增加。在第12周时，骨刻灵组的BMD和骨矿盐含量分别比阴性对照组高4.5%和20.6%。此外，体重顺序为阳性对照组>骨刻灵组>阴性对照组，各组间差异有统计学意义（P<0.05）。股骨切片的显微CT分析显示，骨刻灵组的骨表面/体积比显著高于阴性对照组。X射线图像显示骨刻灵组有清晰的骨痂。此外，高电压显微CT显示骨刻灵组有大量皮质骨堆积。骨刻灵组骨痂的灰度值高于阴性组。从第4周到12周，骨刻灵组血清骨碱性磷酸酶水平升高49.6%，血清Ⅰ型前胶原前肽水平降低80.6%。茜素红染色显示骨刻灵组的钙沉积高于阴性对照组。值得注意的是，骨刻灵组中关键的成骨抑制因子Mgp的表达低于阴性对照组。此外，通过激活核因子κB配体的转化生长因子受体激活剂途径，骨刻灵组中骨形态发生蛋白-2和骨钙素相关蛋白（Bglap）的表达更高。