A network pharmacology approach and validation experiments to investigate the mechanism of the Chinese medicine Wen-Dan decoction in the treatment of steroid-induced femoral head necrosis

Abstract Objectives: The present work studied the mechanism of action of Wen-Dan decoction in treating steroid-induced necrosis of the femoral head (SINFH) via network pharmacology and in vivo experiments. Methods: The active compounds of Wen-Dan decoction and SINFH-related target genes were identified through public databases. Based on the results, further network pharmacological analysis was conducted to explore the potential key active compounds, core targets and biological processes of Wen-Dan decoction in SINFH. Subsequently, the potential mechanisms of action of Wen-Dan decoction in the treatment of SINFH obtained by network pharmacology analyses were validated through in vivo experiments. Results: A total of 608 DEGs (230 upregulated and 378 downregulated) in SINFH were identified. A GO functional enrichment analysis revealed that the SINFH-related genes were mainly involved in neutrophil activation and the immune response, and a KEGG pathway analysis showed that the SINFH-related genes were mainly associated with cytokine receptor interactions, lipids and atherosclerosis, and tuberculosis. In addition, a total of 147 active ingredients of Wen-Dan decoction were identified; of these, the core ingredient was quercetin, and licorice was an active ingredient. Moreover, 277 target genes in the treatment of SINFH with Wen-Dan decoction were identified, and NCF1, PTGS2, and RUNX2 were selected as core target genes. A quantitative real-time polymerase chain reaction (qRT–PCR) assay of peripheral blood from SINFH patients and control individuals showed that the patients exhibited highest levels of PGTS2 and NCF1 in blood. The qRT–PCR analysis of peripheral blood and femoral bone tissue from a mouse model of SINFH and control mice treated with quercetin showed that the PGTS2 and NCF1 levels revealed lower levels in the experimental animals compared with the controls, which was consistent with the bioinformatics analysis results. HE staining, immunohistochemistry and TUNEL staining confirmed significant improvement in hormone-induced femoral head necrosis in the mice treated with quercetin. Conclusions: This study provides new insights into the genes and related pathways involved in SINFH and shows that PTGS2 and NCF1 could be used as potential drug targets in SINFH. Quercetin could improve SINFH by promoting osteogenesis and inhibiting apoptosis.