Objective

2

To explore the possible mechanism of

Huangqi Guizhi Wuwu Decoction

（黄芪桂枝五物汤， HGWD） and

Si Shen Decoction

（SSD） in treating rheumatoid arthritis （RA）.

Methods

2

The disease targets of RA and the herbal medicine components targets of HGWD and SSD， asnd the information on channel entry and syndromes were collected using network pharmacology. Cytoscape 3.9.0 software was used to construct herbal medicine-components-targets-disease network， herbal medicine-channel entry network， and herbal medicine-syndromes network. The protein interaction network was constructed based on the STRING database， and the KEGG pathway enrichment analysis was performed. The active components were selected using the “herbal medicine-ingredients-targets-disease” network， and the core targets were screened out by the protein interaction network. Molecular docking was conducted based on the components and the targets. Experimental cell research was performed for verification. Fibroblast-like synoviocytes （FLS） was induced by 10 ng/ml interleukin-1β （IL-1β） to establish a cellular inflammation model， and then different concentrations of HGWD， SSD and their disassembled formulas were administered for 24 hours. The survival rate of FLS cells and the level of IL-6 were detected by MTT assay and ELISA method. The RAW264.7 cell inflammation model was established， and the active components selected through the “herbal medicine-ingredients-targets-disease” network were administered for 24 hours， after which the nitric oxide （NO） content in the cell supernatant and the expression of P53 gene in the cells were detected.

Results

2

The main active components of HGWD and SSD in the treatment of rheumatoid arthritis included quercetin， kaempferol， isorhamnetin， formononetin， gallic acid， catechin， Taxifolin， luteolin， baicalein， and wogonin through drug target network analysis. According to the protein interaction analysis， the core targets including AKT1， PIK3CA， HRAS， NCOA1， TP53， STAT3， and CCNA2 were obtained， of which TP53 target had the strongest affinity with ten components. In addition， enrichment analysis showed that HGWD and SSD exerted their actions by regulating various signaling pathways such as PI3K-AKT， MAPK， and JAK/STAT. Experimental cell research found that HGWD， SSD and their disassembled formulas at different concentrations could inhibit the inflammatory proliferation and the secretion of IL-6 in FLS cells （

P

＜0.01）. Except for catechin， the other active components could inhibit NO secretion in the supernatant of RAW264.7 cells， and additionally Formononetin could promote the expression of P53 gene （

P

＜0.01）.

Conclusion

2

HGWD， with compositions of Guizhi （

Ramulus Cinnamomi

） and Baishao （

Radix Paeoniae Alba

） assisting Huangqi （

Radix Astragali

）， exerts its function by acting on core targets such as AKT1， PIK3CA， and TP53 to regulate PI3K/Akt and MAPK signaling pathways in the treatment of RA. SSD focuses on restricting the warming and supplementing function of Huangqi by using Jinyinhua （

Flos Lonicerae Japonicae

）， Niuxi （

Radix Achyranthis Bidentatae

） and Shihu （

Caulis Dendrobii

）， and plays its role of treating RA by acting on core targets such as AKT1， STAT3， and TP53 to regulate PI3K-AKT and JAK/STAT signaling pathways. Formononetin may treat RA by adjusting the TP53 target.