Exploring the progress of the traditional Chinese medicine "Duhuo-Qianghuo" in targeting the PI3K-AKT pathway to alleviate knee osteoarthritis based on bioinformatics

This study aims to explore how the traditional Chinese medicine " Du huo-Qiang huo " targets the PI3K-AKT signaling pathway to alleviate knee osteoarthritis using network pharmacology, molecular docking technology, and experimental validation. Active ingredients and target genes related to " Du huo-Qiang huo " and knee osteoarthritis were retrieved from relevant databases.Overlapping data were used to develop a protein-protein interaction (PPI) network model. Functional enrichment analyses, including Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, were performed using the DAVID tool. The identification of core active components and target genes was based on their interaction intensity within the PPI network.These identified components and targets were subjected to molecular docking analysis via AutoDockTools-1.5.7.Furthermore, the therapeutic effects of " Du huo-Qiang huo " in reducing cartilage degradation and inhibiting inflammatory responses were assessed using a knee osteoarthritis mouse model. " Du huo-Qiang huo " comprises 65 bioactive components and 622 associated targets. Through the integration of several databases, 5215 target genes related to knee osteoarthritis were identified, among which 304 were shared between datasets. The primary active compounds implicated in the therapeutic effects of " Du huo-Qiang huo " include homostephanoline, xanthoxyletin , a ngelicone , and ostruthin . Critical therapeutic targets encompass tumor necrosis factor (TNF), protein kinase B1 (AKT1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), interleukin-1β (IL-1β), and albumin (ALB). GO functional enrichment analysis revealed 1231 results, comprising 917 biological processes, 106 cellular components, and 208 molecular functions.KEGG pathway enrichment analysis identified 176 pathways, with the top 20 ranked by P-values and presented in bubble plots. Molecular docking studies demonstrated robust binding affinities between homostephanoline, xanthoxyletin , and targets such as AKT1 and TNF. Additionally, " Du huo-Qiang huo " granules showed partial restorative effects on cartilage damage and a reduction in inflammatory markers. This study offers a preliminary exploration of the mechanisms underlying the therapeutic effects of " Du huo-Qiang huo " in managing knee osteoarthritis. Through an analysis of its diverse components, molecular targets, and related pathways, the findings highlight that pivotal active compounds, such as homostephanoline and xanthoxyletin , engage with critical targets like TNF and AKT1.These molecular interactions modulate the PI3K-AKT signaling pathway, promoting cartilage regeneration and mitigating inflammatory responses,which collectively contribute to its therapeutic efficacy in knee osteoarthritis.