Network pharmacology and experimental validation to identify the potential mechanism of Hedyotis diffusa Willd against rheumatoid arthritis

Rheumatoid arthritis (RA) is a chronic, systemic, autoimmune disease that may lead to joint damage, deformity, and disability, if not treated effectively. Hedyotis diffusa Willd (HDW) and its main components have been widely used to treat a variety of tumors and inflammatory diseases. The present study utilized a network pharmacology approach, microarray data analysis and molecular docking to predict the key active ingredients and mechanisms of HDW against RA. Eleven active ingredients in HDW and 180 potential anti-RA targets were identified. The ingredients-targets-RA network showed that stigmasterol, beta-sitosterol, quercetin, kaempferol, and 2-methoxy-3-methyl-9,10-anthraquinone were key components for RA treatment. KEGG pathway results revealed that the 180 potential targets were inflammatory-related pathways with predominant enrichment of the AGE-RAGE, TNF, IL17, and PI3K-Akt signaling pathways. Screened through the PPI network and with Cytoscape software, RELA, TNF, IL6, TP53, MAPK1, AKT1, IL10, and ESR1 were identified as the hub targets in the HDW for RA treatment. Molecular docking was used to identify the binding of 5 key components and the 8 related-RA hub targets. Moreover, the results of network pharmacology were verified by vitro experiments. HDW inhibits cell proliferation in MH7A cells in a dose and time-dependent manner. RT-qPCR and WB results suggest that HDW may affect hub targets through PI3K/AKT signaling pathway, thereby exerting anti-RA effect. This study provides evidence for a clinical effect of HDW on RA and a research basis for further investigation into the active ingredients and mechanisms of HDW against RA.

Automated summary

In this study, a network pharmacology approach, microarray data analysis, and molecular docking were used to predict the key active ingredients and mechanisms of Hedyotis diffusa Willd (HDW) against rheumatoid arthritis (RA). The results showed that HDW contains 11 active ingredients and 180 potential anti-RA targets. The study also identified stigmasterol, beta-sitosterol, quercetin, kaempferol, and 2-methoxy-3-methyl-9,10-anthraquinone as key components for RA treatment. Pathway analysis revealed that the potential targets were mainly related to inflammatory pathways such as AGE-RAGE, TNF, IL17, and PI3K-Akt signaling pathways. Furthermore, RELA, TNF, IL6, TP53, MAPK1, AKT1, IL10, and ESR1 were identified as hub targets for HDW in treating RA. Molecular docking confirmed the binding of the key components and the hub targets. In vitro experiments demonstrated that HDW inhibits cell proliferation in MH7A cells and affects hub targets through the PI3K/AKT signaling pathway. This study provides evidence for the clinical effect of HDW on RA and lays a foundation for further investigation into its active ingredients and mechanisms against RA.