Predicting and Exploring the Mechanisms of Erzhi Pill in Prevention and Treatment of Osteoporosis Based on Network Pharmacology and Zebrafish Experiments

Abstract

Background

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.

Methods

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.

Results

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.

Conclusion

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.

Keywords:

• Erzhi Pill
• osteoporosis
• network pharmacology
• mechanisms
• zebrafish
• quercetin

Acknowledgments

The authors are thankful to the zebrafish platform of clinical research center, Affiliated Hospital of Guangdong Medical University for use of the zebrafish.

Data Sharing Statement

The data that support the findings of the present study are available in Supplementary Materials.

Author Contributions

All authors made a significant contribution to the work reported, whether that is in the conception, study design, execution, acquisition of data, analysis and interpretation, or in all these areas; took part in drafting, revising or critically reviewing the article; gave final approval of the version to be published; have agreed on the journal to which the article has been submitted; and agree to be accountable for all aspects of the work. Zhiguo Zhong and Yuyun Li are the co-first authors.

Disclosure

All authors declare that they have no conflicts of interest for this work.

Additional information

Funding

The study was supported by the Natural Science Foundation of Guangdong Province (No: 2018A030313390), the Scientific Research Foundation for PhD of Guangdong Medical University (No: B2017030), and the Scientific Research Subject of Traditional Chinese Medicine Bureau of Guangdong Province (No: 20171145 & 20191190).