Mechanism of Tongbi Jiangu Prescription in Treatment of Knee Osteoarthritis Based on Network Pharmacology and Molecular Docking / 中国实验方剂学杂志

ABSTRACT

ObjectiveTo explore the mechanism of Tongbi Jiangu prescription （TBJG） in the treatment of knee osteoarthritis （KOA） based on network pharmacology and molecular docking，and further verify it by cell experiments. MethodThe active components and the corresponding targets of TBJG were screened out according to the traditional Chinese medicine systems pharmacology database and analysis platform（TCMSP）. The targets of KOA were obtained from GeneCards，online mendelian inheritance in man（OMIM）， and DrugBank. The common targets of active components of TBJG and KOA were the targets of TBJG against KOA. The active component-target network and protein-protein interaction （PPI） network were constructed by Cytoscape 3.7.2. STRING was used for PPI network analysis. DAVID was used for gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） enrichment analyses. Key targets and core active components were selected for molecular docking by AutoDock. The results of network pharmacology were verified by cell experiments and the pharmacodynamic responses were observed. ResultThe prediction of network pharmacology showed that there were 111 active components of TBJG in the treatment of KOA. The core active components were quercetin，kaempferol， and β-sitosterol，and the key targets were interleukin-1β（IL-1β）， matrix metalloproteinase-3 （MMP-3），and tumor necrosis factor-α （TNF-α）. Biological processes （BP） in GO analysis mainly involved inflammatory response，response to lipopolysaccharide，apoptosis signaling pathway，and regulation of DNA activity in binding transcription factor. Cellular components （CC） included plasma membrane protein complex，RNA polymerase Ⅱ transcription factor complex，membrane raft，and serine/threonine protein kinase complex. Molecular functions （MF） were mainly enriched in cytokine receptor binding，nuclear receptor activity，protein domain specific binding，serine hydrolase activity，chemokine receptor binding，and activity of nitric oxide synthase regulator. As revealed by the KEGG analysis， the relevant signaling pathways were nuclear factor（NF）-κB， Janus kinase（JAK）/signal transducer and activator of transcription（STAT）， and Wnt signaling pathways. Molecular docking results showed that the core active components had good binding activities with key targets. The experimental results showed that TBJG could down-regulate IL-1β， MMP-3，TNF-α， and NF-κB p65 expression levels （P<0.05），and up-regulated NF-κB inhibitor（IκB）-α（P<0.05）. ConclusionThe mechanism of TBJG in the treatment of KOA lies in the application of active components such as quercetin，kaempferol， and β-sitosterol with IL-1β，MMP-3， and TNF-α as key targets through the NF-κB，JAK/STAT， and Wnt signaling pathways.