A synthetic peptide from Sipunculus nudus promotes bone formation via Estrogen/MAPK signal pathway based on network pharmacology

Peiran Wang; Zhenhui Feng; Siyu Chen; Yingye Liang; Haiyan Hou; Qianqian Ouyang; Hui Yu; Hua Ye; Lei Cai; Yi Qi; Kefeng Wu; Hui Luo

doi:10.3389/fphar.2023.1173110

A synthetic peptide from Sipunculus nudus promotes bone formation via Estrogen/MAPK signal pathway based on network pharmacology

Front Pharmacol. 2023 Apr 24:14:1173110. doi: 10.3389/fphar.2023.1173110. eCollection 2023.

Authors

Peiran Wang¹, Zhenhui Feng^{1

2}, Siyu Chen^{1

2}, Yingye Liang^{1

2}, Haiyan Hou¹, Qianqian Ouyang^{1

2

3}, Hui Yu^{1

2

4}, Hua Ye^{1

2

3}, Lei Cai⁵, Yi Qi^{1

2

3}, Kefeng Wu^{1

2

3}, Hui Luo^{1

2

3}

Affiliations

¹ Marine Biomedical Research Institution, Guangdong Medical University, Zhanjiang, China.
² Guangdong (Zhanjiang) Provincial Laboratory of Southern Marine Science and Engineering, Zhanjiang, China.
³ Marine Traditional Chinese Medicine Sub-center of National Engineering Research Center for Modernization of Traditional Chinese Medicine, Zhanjiang, China.
⁴ Guangdong Key Laboratory for Research and Development of Natural Drugs, Guangdong Medical University, Zhanjiang, China.
⁵ Guangdong Provincial Key Laboratory of Laboratory Animals, Guangdong Laboratory Animals Monitoring Institute, Guangzhou, China.

Abstract

The tripeptide Leu-Pro-Lys (LPK), derived from the Sipunculus nudus protein, was synthesized and studied to investigate its potential protective effect on bone formation. The effect and mechanism of LPK were analyzed through network pharmacology, bioinformatics, and experimental pharmacology. The study found that LPK at concentrations of 25 μg/mL and 50 μg/mL significantly increased ALP activity and mineralization in C3H10 cells. LPK also increased the expression of COL1A1 and promoted bone formation in zebrafish larvae. Network pharmacology predicted 148 interaction targets between LPK and bone development, and analysis of the protein-protein interaction network identified 13 hub genes, including ESR1, MAPK8, and EGFR, involved in bone development. Through KEGG enrichment pathways analysis, it was determined that LPK promotes bone development by regulating endocrine resistance, the relaxin signaling pathway, and the estrogen signaling pathway. Molecular docking results showed direct interactions between LPK and ESR1, MAPK8, and MAPK14. Additional verification experiments using western blot assay revealed that LPK significantly upregulated the expression of genes related to bone formation, including COL1A1, OPG, RUNX2, ESR1, phosphorylated MAPK14, and phosphorylated MAPK8 in C3H10 cells. These results suggest that LPK promotes bone formation by activating the estrogen/MAPK signaling pathway.

Keywords: MAPK signaling pathway; bone development; estrogen signaling pathway; network pharmacology; sipunculus nudus; zebrafish.

Grants and funding

This work was supported by the Fund of Southern Marine Science and Engineering Guangdong Laboratory (Zhanjiang) (ZJW-2019-007), the Science and Technology Program of Guangdong Province (2019B090905011, 2018A030307001), the Special Funds for Economic Development of Marine Economy of Guangdong Province, China (GDME-2018C011), the Special Science and Technology Innovation Project of Guangdong Province, China (2022A01207, 2019A03023), the Discipline Construction Project of Guangdong Medical University, China (4SG22009G, 4SG22264G), and the Ph.D. Start-up Fund of Guangdong Medical University (GDMUB2022025). The authors acknowledge the technical support of the Public Service Platform for Research and Development of Marine Biomedical Resources in the South China Sea.