Sun Weipeng, Li Minying, Xie Lei, Mai Zhexing, Zhang Yan, Luo Lieliang, Yan Zijian, Li Zige, Dong Hang, Huang Feng, Shen Zhen, Jiang Ziwei
The First School of Clinical Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong Province, China.
Medical College of Acupuncture-Moxibustion and Rehabilitation, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong Province, China.
Front Pharmacol. 2021 May 31;12:603734. doi: 10.3389/fphar.2021.603734. eCollection 2021.
Drynariae Rhizoma (DR) has been demonstrated to be effective in promoting fracture healing in clinical use. In the study, we tried to predicate potential signaling pathways and active ingredients of DR network pharmacology, uncover its regulation mechanism to improve large bone defects by and experiment. We total discovered 18 potential active ingredients such as flavonoids and 81 corresponding targets, in which mitogen-activated protein kinase (MAPK) signaling pathway has the highest correlation with bone defects in pathway and functional enrichment analysis. Therefore, we hypothesized that flavonoids in DR improve large bone defects by activating MAPK signaling pathway. Animal experiments were carried out and all rats randomly divided into TFDR low, medium, and high dosage group, model group and control group. 12 weeks after treatment, according to X-ray and Micro-CT, TFDR medium dosage group significantly promote new bone mineralization compared with other groups. The results of HE and Masson staining and in vitro ALP level of BMSC also demonstrated the formation of bone matrix and mineralization in the TFDR groups. Also, angiographic imaging suggested that flavonoids in DR promoting angiogenesis in the defect area. Consistently, TFDR significantly enhanced the expression of BMP-2, RUNX-2, VEGF, HIF-1 in large bone defect rats based on ELISA and Real-Time PCR. Overall, we not only discover the active ingredients of DR in this study, but also explained how flavonoids in DR regulating MAPK signaling pathway to improve large bone defects.
骨碎补在临床应用中已被证明对促进骨折愈合有效。在本研究中，我们尝试通过网络药理学预测骨碎补的潜在信号通路和活性成分，通过实验揭示其改善大骨缺损的调控机制。我们共发现了18种潜在活性成分，如黄酮类化合物，以及81个相应靶点，其中丝裂原活化蛋白激酶（MAPK）信号通路在通路和功能富集分析中与骨缺损的相关性最高。因此，我们推测骨碎补中的黄酮类化合物通过激活MAPK信号通路改善大骨缺损。进行了动物实验，所有大鼠随机分为骨碎补总黄酮低、中、高剂量组、模型组和对照组。治疗12周后，根据X射线和显微CT检查，骨碎补总黄酮中剂量组与其他组相比显著促进了新骨矿化。HE和Masson染色结果以及骨髓间充质干细胞的体外碱性磷酸酶水平也证明了骨碎补总黄酮组有骨基质形成和矿化。此外，血管造影成像表明骨碎补中的黄酮类化合物促进了缺损区域的血管生成。同样，基于ELISA和实时定量PCR，骨碎补总黄酮显著增强了大骨缺损大鼠中骨形态发生蛋白-2（BMP-2）、 Runt相关转录因子2（RUNX-2）、血管内皮生长因子（VEGF）、缺氧诱导因子-1（HIF-1）的表达。总体而言，我们不仅在本研究中发现了骨碎补的活性成分，还解释了骨碎补中的黄酮类化合物如何调节MAPK信号通路以改善大骨缺损。
