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| Exploration of the mechanism of Bushen Bitong Prescription in the treatment of knee osteoarthritis based on network pharmacology and experimental verification |
| XIANG Wenyuan1 YI Lin1 DENG Yingjie2 FANG Rui2▲ |
1.The Fourth School of Clinical Medicine, Xinjiang Medical University, Xinjiang Uygur Autonomous Region, Urumqi 830054, China;
2.the Second Department of Orthopaedics, Hospital of Traditional Chinese Medicine, Xinjiang Medical University, Xinjiang Uygur Autonomous Region, Urumqi 830099, China
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Abstract Objective To explore the mechanism of Bushen Bitong Prescription in the treatment of knee osteoarthritis (KOA) by network pharmacology and experimental verification. Methods Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform were used to obtain the active ingredients and targets of Bushen Bitong Prescription. GeneCards, OMIM, PharmGkb, TTD, and DrugBank databases were used to search KOA disease related targets, and the intersection of drug and disease targets was selected. The obtained active ingredients and related targets were introduced into Cytoscape software, and the network map of “Bushen Bitong Prescription - active ingredients-KOA targets” was drawn to screen the key active ingredients. Potential targets were imported into the STRING database to plot the protein-protein interaction network. The core targets of Bushen Bitong Prescription in the treatment of KOA were screened, and GO enrichment and KEGG annotation were analyzed. Human chondrocytes were selected and set up in control group, injury group, and administration group. The control group was cultured normally, the injury group and administration group were cultured in the medium containing interleukin (IL)-1β (10 ng/ml) DMEM /F12, and the administration group was treated with 0.444 7 μg/ml Bushen Bitong Prescription liquid. After 24 h of culture, the cell morphology of the three groups was observed. The levels of inflammatory factors were detected by enzyme-linked immunosorbent assay, and the expression of cartilage repair factors in the three groups was detected by qRT-PCR and Western blot. Results A total of 186 active ingredients of Bushen Bitong Prescription were screened, mainly including quercetin, luteolin, kaempferol, etc., mainly through the key targets of TNF, MMP1, CXCL2, IL1B, and signaling pathways of PI3K/Akt, T cell receptors, toll-like receptors, hypoxy-inducer factor-1, TNF, IL-17, and others. It involved, oxidative stress, apoptosis, inflammation, and other biological processes. Microscopically, the number of cells decreased in the injury group and increased in the administration group. The levels of tumor necrosis factor-α (TNF-α), IL-1β, and IL-6 in injury group were higher than those in control group, and the levels of transforming growth factor-β (TGF-β), insulin-like growth factor-1 (IGF-1), basic fibroblast growth factor (BFGF), and the gene and protein expression levels of SOX9 and COL2A1 were lower than those in control group (P＜0.05). The levels of TNF-α, IL-1β, and IL-6 in administration group were lower than those in injury group, and the levels of TGF-β, IGF-1, BFGF, and the gene and protein expression levels of SOX9 and COL2A1 were higher than those in injury group (P＜0.05). Conclusion Bushen Bitong Prescription may promote the repair of injured chondrocytes by relieving inflammatory cell infiltration and participating in the regulation of SOX9 and COL2A1.
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