Mechanism Investigation of Wuwei Shexiang Pills on Gouty Arthritis via Network Pharmacology, Molecule Docking, and Pharmacological Verification

Abstract

Background. Gout is a common crystal-related arthritis caused by the deposition of monosodium urates (MSU). Tibetan medicine Wuwei Shexiang Pills (WSP) has been demonstrated to exhibit anti-inflammatory, antihyperuricemia, and antigout activities. However, the underlying mechanism is unknown. Objectives. To explore the mechanisms of Wuwei Shexiang Pills on gouty arthritis via network pharmacology, molecule docking, and pharmacological verification. Methods. The ingredients and targets of WSP were obtained by searching and screening in BATMAN-TCM and SwissADME. The targets involving the gout were acquired from public databases. The shared targets were put onto STRING to construct a PPI network. Furthermore, Metascape was applied for the GO and KEGG enrichment analysis to predict the biological processes and signaling pathways. And molecular docking was performed to validate the binding association between the key ingredients and the relative proteins of TNF signaling. Based on the serum pharmacology, the predicted antigout mechanism of WSP was validated in MSU-induced THP-1 macrophages. The levels of inflammatory cytokines and mRNA were measured by ELISA and qRT-PCR, respectively, and MAPK, NF-κB, and NLRP3 signaling-associated proteins were determined by western blot and immunofluorescence staining. Results. 48 bioactive ingredients and 165 common targets were found in WSP. The data showed that 5-Cis-Cyclopentadecen-1-One, 5-Cis-Cyclotetradecen-1-One, (−)-isoshyobunone, etc. were potential active ingredients. TNF signaling, HIF-1 signaling, and Jak-STAT signaling were predicted to be the potential pathways against gout. The molecule docking analysis found that most ingredients had a high affinity for p65, NLRP3, IL-1β, TNF-α, and p38. The data from in vitro experiment showed that WSP suppressed the production and gene expression of inflammatory cytokines. Furthermore, WSP could inhibit the activation of MAPK, NF-κB, and NLRP3 signaling pathways. Conclusion. Our finding suggested that the antigout effect of WSP could be achieved by inhibiting MAPK, NF-κB, and NLRP3 signaling pathways. WSP might be a candidate drug for gouty treatment.

LC-MS-based metabolomics reveals the mechanism of anti-gouty arthritis effect of Wuwei Shexiang pill

“…Recently, WSP has also been reported to significantly reduce serum uric acid levels and inhibit ear swelling in mice ( Yamin and Lvyi, 2017 ). In our previous study, we confirmed that WSP exerted an anti-gout effect by inhibiting MAPK, NF-κB, and NLRP3 signaling pathways in MSU-stimulated THP-1 macrophages ( Lang et al, 2022 ). Although our research has unveiled the anti-inflammatory effect and mechanism in vitro , the anti-gout pharmacodynamic activity and the metabolomic profiles of WSP in vivo have not been reported.…”

“…WSP has been used in China, especially among Tibetan people, for the treatment of inflammation and inflammation-related diseases, such as tonsillitis, pharyngitis, and rheumatoid arthritis. Our preliminary study found that WSP might exert an anti-gout effect through multiple targets and pathways ( Lang et al, 2022 ). However, the specific mechanism is unclear.…”

“…Thus, key indicators of the linoleic acid metabolic pathway, such as lipoxygenase and cytochrome P4501A2, could be subsequently validated by qRT-PCR, Western blot, and ELISA technology ( Wang et al, 2022 ). In consideration of the inhibition of WSP on the MAPK/NF-κB signaling pathway ( Lang et al, 2022 ), we hypothesized that WSP inhibited the combination of NF-κB and target DNA, thereby suppressing COX expression and further reducing the production of related metabolites catalyzed by COX1/2, such as PGs. In the present study, we elucidated the role of WSP in the NF-κB signaling pathway from a metabolomics perspective for the first time.…”

Anti-Hyperuricemic, Anti-Arthritic, Hemolytic Activity and Therapeutic Safety of Glycoconjugated Triazole-Phthalimides

“…The therapy of hyperuricemia and the severity of gouty arthritis still present challenges, mainly due to adverse reactions and toxic effects of commercial therapeutic agents [ 23 , 24 ]. Therefore, the search for molecules that are efficient in reducing the serum level of uric acid in the hyperuricemic state and decreasing urates and glycoproteins and the inflammatory process of gouty arthritis is fundamental and urgent [ 25 , 26 ]. In this scenario, phthalimides are a group of cyclic imides with diverse biological activities, from hypolipidemic [ 27 ] to analgesic, antitumor, antimicrobial [ 28 ], antifungal [ 29 ], antiparasitic [ 30 ], antiviral for the treatment and/or prophylaxis of COVID-19 [ 31 ], anticonvulsant, antipyretic [ 32 ] and anti-inflammatory [ 33 ], as well as the recently reported antihyperlipidemic activity of glycoconjugated phthalimides in mice subjected to a model of dyslipidemia and insulin resistance [ 34 ].…”