Network pharmacology and molecular docking analysis on mechanism of osthole in treating knee osteoarthritis
10.3969/j.issn.1000-4718.2024.12.014
- VernacularTitle:基于网络药理学和分子对接探讨蛇床子素治疗膝骨关节炎的机制
- Author:
Haoyu WANG
1
;
Aidi LIANG
;
Wei YAO
;
Xiaoyun LI
;
Zhuo HUANG
;
Rong-hua ZHANG
Author Information
1. 暨南大学中医学院,广东 广州 510632
- Publication Type:Journal Article
- Keywords:
osthole;
osteoarthritis;
chondrocyte;
interleukin-1β;
inflammation
- From:
Chinese Journal of Pathophysiology
2024;40(12):2302-2311
- CountryChina
- Language:Chinese
-
Abstract:
AIM:This study aims to investigate the effects of Osthole(OST)on inflammation and cartilage-de-grading proteases in an interleukin 1β(IL-1β)-induced chondrocyte inflammation model.METHODS:Prediction:we collected target genes for OST and those related to knee osteoarthritis(KOA)from four databases.After standardizing the target genes obtained from the UniProt database,two overlapping genes were identified using the Venny tool.Additional-ly,protein interaction relationships were obtained from the STRING database,and core target genes were screened and vi-sualized using Cytoscape software.Enrichment analysis for the overlapping genes was performed through Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways.Finally,the active drug components and target proteins were validated and visualized through computational analysis.Validation:primary rabbit chondrocytes were iso-lated and cultured in vitro.Cell morphology and toluidine blue staining were employed to confirm chondrocyte identity.An inflammatory injury model was induced using IL-1β.The cells were divided into control,model,low-dose,medium-dose,high-dose OST,and celecoxib groups.Chondrocyte viability was assessed using the CCK-8 method.Western blot and im-munofluorescence techniques were utilized to detect the proteins IL-1β,tumor necrosis factor-alpha(TNF-α),matrix me-talloproteinase 13(MMP-13),and a disintegrin and metalloproteinase with thrombospondin motifs(ADAMTS-4).Gene expression levels of IL-1β,IL-6,TNF-α,and MMP-13 were measured via RT-qPCR.RESULTS:A total of 80 potential OST targets were identified for treating KOA,with 10 core target genes(CASP3,TNF,HIF1A,IL-1β,NFKB1,PARP1,NFE2L2,JAK2,MAPK1,and GSK3B)screened.GO and KEGG analyses further elucidated the molecular mechanisms of OST in KOA treatment,highlighting multiple biological processes and signaling pathways involved.Molecular docking simulations confirmed stable binding of OST to key targets TNF and IL-1β within the IL-17 signaling pathway.Chondro-cytes exhibited long spindle and pavement-like shapes.Based on the effects of varying OST concentrations on chondro-cytes,2.5,5,and 10 μmol/L OST were selected for pharmacodynamic testing.Compared to the model group,OST signif-icantly reduced inflammation in the chondrocyte inflammation model and inhibited the expression of cartilage matrix-de-grading enzymes,showing no statistically significant difference from the celecoxib group.CONCLUSION:OST promotes chondrocyte proliferation and differentiation.It effectively inhibits inflammation in the IL-1β-induced chondrocyte model and mitigates chondrocyte injury.The underlying mechanism may involve the degradation of chondroproteoglycans and the protection of the extracellular matrix.
