Research Paper Volume 14, Issue 14 pp 5727—5748

The therapeutic effect of Fufang Zhenshu Tiaozhi (FTZ) on osteoclastogenesis and ovariectomized-induced bone loss: evidence from network pharmacology, molecular docking and experimental validation

Xiaojun Chen1, *, , Jiangyan Wang2, *, , Lin Tang2, , Qiuying Ye3, , Qunwei Dong4,5, , Zhangwei Li6, , Li Hu2, , Chenghong Ma2, , Jiake Xu7, , Ping Sun2, ,

  • 1 School of Molecular Sciences, University of Western Australia, Perth 6009, Western Australia, Australia
  • 2 Department of Endocrinology, The First Affiliated Hospital of Guangdong Pharmaceutical University, Guangzhou 510000, Guangdong, China
  • 3 College of Food and Medicine, Qingyuan Polytechnic, Qingyuan 511510, Guangdong, China
  • 4 Department of Orthopedic, The First Affiliated Hospital of Guangdong Pharmaceutical University, Guangzhou 510000, Guangdong, China
  • 5 Department of Orthopedic, Yunfu Hospital of Traditional Chinese Medicine, Yunfu 527300, Guangdong, China
  • 6 Department of Stomatology, The First Affiliated Hospital of Guangdong Pharmaceutical University, Guangzhou 510000, Guangdong, China
  • 7 School of Biomedical Sciences, University of Western Australia, Perth 6009, Western Australia, Australia
* Equal contribution

Received: February 21, 2022       Accepted: June 23, 2022       Published: July 12, 2022      

https://doi.org/10.18632/aging.204172
How to Cite

Copyright: © 2022 Chen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Fufang Zhenshu Tiaozhi (FTZ) has been widely used in clinical practice and proven to be effective against aging-induced osteoporosis in mice. This study aimed to explore the mechanism of FTZ against osteoclastogenesis and ovariectomized-induced (OVX) bone loss through the network pharmacology approach. The ingredients of FTZ were collected from the previous UPLC results, and their putative targets were obtained through multiple databases. Differentially expressed genes (DEGs) during osteoclastogenesis were identified through multi-microarrays analysis. The common genes between FTZ targets and DEGs were used to perform enrichment analyses through the clusterProfier package. The affinity between all FTZ compounds and enriched genes was validated by molecular docking. The effects of FTZ on osteoclastogenesis and bone resorption were evaluated by TRAP staining, bone resorption assay and RT-qPCR in vitro, while its effects on bone loss by ELISA and Micro-CT in vivo. Enrichment analyses indicated that the inhibitory effects of FTZ may primarily involve the regulation of inflammation, osteoclastogenesis, as well as TNF-α signaling pathway. 130 pairs docking results confirmed FTZ ingredients have good binding activities with TNF-α pathway enriched genes. FTZ treatment significantly reduced TRAP, TNF-α, IL-6 serum levels and increased bone volume in OVX mice. Consistently, in vitro experiments revealed that FTZ-containing serum significantly inhibited osteoclast differentiation, bone resorption, and osteoclast related mRNA expression. This study revealed the candidate targets of FTZ and its potential mechanism in inhibiting osteoclastogenesis and bone loss induced by OVX, which will pave the way for the application of FTZ in the postmenopausal osteoporosis treatment.

Abbreviations

FTZ: Fufang Zhenshu Tiaozhi; UPLC: Ultra-performance liquid chromatography; DEGs: Differentially expressed genes; RRA: Robust rank aggregation; GOBP: Gene Ontology Biological Processes; PPI: Protein-protein interaction; TCMSP: the traditional Chinese medicine system pharmacology database and analysis platform; STP: Swiss Target Prediction platform; SEA: Similarity ensemble approach platform; GEO: Gene Expression Omnibus; OVX: Ovariectomy.