Traditional Chinese Medicine Department, Affiliated Hospital of Guangdong Medical University, Zhanjiang, 524023, People's Republic of China.
Department of Pharmacology, School of Pharmacy, Guangdong Key Laboratory for Research and Development of Natural Drugs, Marine Biomedical Research Institute, Guangdong Medical University, Zhanjiang, 524023, People's Republic of China.
Drug Des Devel Ther. 2021 Feb 24;15:817-827. doi: 10.2147/DDDT.S293455. eCollection 2021.
Erzhi Pill (EZP), a traditional Chinese medicine (TCM) prescription, has been widely applied to improve bone metabolism and treat osteoporosis (OP) in China. However, its effective constituents and mechanisms remain unclear.
By combining network pharmacology and zebrafish experiments, an integrative method was employed to address this problem. Firstly, the disease targets of OP were collected from two public gene databases. Secondly, the active compounds and drug targets of EZP were obtained from the traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP). Thirdly, a drug-target-disease interaction network was constructed, and the key active components were identified by analyzing the topological characteristics of the network. Finally, these predicted results were tested by zebrafish experiments and compared with those from the literature. Specifically, quercetin as an important representative active component of EZP was applied to wild type and transgenic zebrafish larvae to assess its effects on skull mineralization and osteoplastic differentiation.
Our study identified 72 active compounds, 220 targets and 166 signaling pathways probably involved in the prevention and treatment of OP by EZP, wherein quercetin, apigenin, daidzein, luteolin, ursolic acid and kaempferol could be the key compounds, while PI3K-Akt signaling pathway, TNF signaling pathway and IL-17 signaling pathway could be the key signaling pathways. The experiments indicated that quercetin attenuated both the decrease of skull mineralization and the inhibition of skull osteoplastic differentiation in zebrafish larvae trigged by dexamethasone.
Our study not only investigated potentially effective constituents and mechanisms of EZP in the prevention and treatment of OP, but also provided a reference for the in-depth research, development and application of TCM.
二至丸（EZP）是一种中药方剂，在中国被广泛用于改善骨代谢和治疗骨质疏松症（OP）。然而，其有效成分和作用机制尚不清楚。
本研究采用网络药理学和斑马鱼实验相结合的方法，对其进行了研究。首先，从两个公共基因数据库中收集 OP 的疾病靶点。其次，从中药系统药理学数据库和分析平台（TCMSP）中获得 EZP 的活性化合物和药物靶点。然后，构建药物-靶点-疾病相互作用网络，并通过分析网络的拓扑特征来确定关键的活性成分。最后，通过斑马鱼实验对这些预测结果进行了验证，并与文献进行了比较。具体来说，将 EZP 的重要代表性活性成分槲皮素应用于野生型和转基因斑马鱼幼鱼，以评估其对颅骨矿化和成骨分化的影响。
本研究共鉴定出 72 种活性化合物、220 个靶点和 166 条可能与 EZP 防治 OP 相关的信号通路，其中槲皮素、芹菜素、大豆苷元、木犀草素、熊果酸和山奈酚可能是关键化合物，而 PI3K-Akt 信号通路、TNF 信号通路和 IL-17 信号通路可能是关键信号通路。实验表明，槲皮素可减轻地塞米松诱导的斑马鱼幼鱼颅骨矿化减少和颅骨成骨分化抑制。
本研究不仅探讨了 EZP 防治 OP 的潜在有效成分和作用机制，还为中药的深入研究、开发和应用提供了参考。
