Network pharmacology and experimental validation to study the potential mechanism of Tongguanteng injection in regulating apoptosis in osteosarcoma

Sep 21, 2026

Upload by Himbo

This study identifies active components of Tongguanteng injection (TGT) and evaluates its preclinical efficacy and mechanism in osteosarcoma by combining network pharmacology with experimental validation. Network construction integrates active ingredient–disease–target–pathway and protein–protein interaction networks, followed by GO/KEGG enrichment, molecular docking, and SwissADME pharmacokinetic prediction. RT-PCR, proliferation, morphology, apoptosis, MMP assays, and Western blotting confirm effects in 143B and SAOS2 cells, showing TGT induces apoptosis via HIF-1 signaling inhibition and PD-1 checkpoint modulation, and alters key apoptosis markers.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages23

Reading Time58 min

AccessFree to download

Abstract

Objective

Methods

Results

Conclusion

Introduction

Osteosarcoma background and treatment challenges

Wei et al. BMC Complementary Medicine and Therapies (2024) 24:67 [https://doi.org/10.1186/s12906-024-04354-z](https://doi.org/10.1186/s12906-024-04354-z) BMC Complementary Medicine and Therapies RESEARCH Open Access Network pharmacology and experimental validation to study the potential mechanism of Tongguanteng injection in regulating apoptosis in osteosarcoma Lanyi Wei1, Jingjing Meng1, Danfeng Xiang1, Quanjun Yang1, Yangyun Zhou1, Lingyan Xu1, Mengyue Wang2, Junjun Chen1* and Yonglong Han 1* Abstract Objective The main objectives ofthis study were to identify the active components of Tongguanteng injection (TGT) and investigate the preclinical efficacy and mechanism of TGT on osteosarcoma using a combination of network pharmacology and experimental validation. Methods To identify the active constituents and targets ofTGT against osteosarcoma using network pharmacology, we constructed a network consisting of an ’active ingredient-disease-target-pathway’ and a protein–protein interaction (PPI) network. The target organ network was utilized to investigate the distribution of core targets in tissues. Afterwards, the core targets underwent Gene ontology (GO) functional enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. The binding energy between receptors and ligands was compared using molecular docking. In addition, SwissADME was employed to forecast the pharmacokinetic characteristics of the substances. Finally, real-time polymerase chain reaction (RT-PCR), cell proliferation assay, morphological analysis, apoptosis assay, mitochondrial membrane potential (MMP) detection, and Western blotting were utilized to confirm the potential mechanisms ofTGT treatment in osteosarcoma cell lines 143B and SAOS2 . Results A total of 54 chemical constituents ofTGT and 71 targets associated with osteosarcoma were acquired. Through the molecular docking technology, Tenacigenin B, Marsdekoiside, Taraxasterol, Tenacissoside G, Tenacissoside L, and Tenacissoside J were identified as the primary active components ofTGT among the various compounds. Analysis of target organs suggests that TGT may play an anti-osteosarcoma role through immune regulation. The GO and KEGG enrichment analysis revealed that TGT could trigger osteosarcoma cell apoptosis by inhibiting the HIF-1 signalling pathway and modulating PD-1 expression and the PD-1 checkpoint pathway in cancer. SwissADME database predicted that Tenacigenin B and Taraxasterol had the best drug-likeness. In vitro studies also demonstrated that TGT suppressed the activity and induced alterations in the morphology of osteosarcoma cells. It decreased MMP levels, triggered apoptosis by increasing Bax expression and Caspase-3 activity, and decreased Bcl-2 expression, thereby exerting an anti-osteosarcoma effect. In the meantime, RT-PCR tests demonstrated that TGT could control immune *Correspondence: Junjun Chen[chenjunjun8812@sina.com](chenjunjun8812@sina.com)[ ](chenjunjun8812@sina.com)Yonglong Han [yonglongh@126.com](yonglongh@126.com) Full list of author information is available at the end of the article © The Author(s) 2024. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit [http://creativecommons.org/licens

FAQs

What is the main purpose of this study on Tongguanteng injection (TGT)?

The study aims to identify TGT’s active components and investigate its preclinical efficacy and underlying mechanism in osteosarcoma by combining network pharmacology and experimental validation.

How were the study targets and pathways predicted?

Network pharmacology generated an active ingredient–disease–target–pathway network and a protein–protein interaction network, followed by target organ distribution analysis, GO and KEGG enrichment, molecular docking, and SwissADME prediction for drug-likeness and pharmacokinetics.

What experimental evidence supports TGT’s mechanism in osteosarcoma cells?

In 143B and SAOS2 cells, TGT suppressed tumor cell activity and morphology changes, reduced mitochondrial membrane potential, increased apoptosis as shown by Bax expression and caspase-3 activity, decreased Bcl-2, and modulated pathways including HIF-1 signaling and PD-1 expression.

Sep 21, 2026

Upload by Himbo