To investigate the potential mechanism of action of Dew-removing and Pain-Relieving Powder (CSDT) in treating gouty arthritis (GA) using network pharmacology and molecular docking techniques.

Methods: (1) Active ingredients, targets, and GA-related targets of Dew-removing and Pain-Relieving Powder were screened from TCMSP, DisGeNET, GeneCards, and OMIM databases. The "drug-component-target-disease" network and protein-protein interaction network were constructed using Cytoseape_v 3.7.1 software, followed by bioinformatics enrichment analysis with DAVID and molecular docking validation with Autodoek. Experimental validation was conducted using AGA mouse models: Forty-eight Kunming mice were randomly divided into normal control group, model control group, colchicine tablet 5 mg/kg group, and CSDT low, medium, and high dose groups (0.975 g/ml, 1.95 g/ml, and 3.9 g/ml), with 8 mice in each group. Except for the blank control group and model group, all mice received pre-gastric administration for 3 days. Acute GA models were established on day 4 after gastric administration, with ankle joint swelling rates and thermal pain thresholds measured at time points (0, 2, 12,24,48, and 72 h) post-modeling. After 3 days of administration, samples were collected for HE staining to observe synovial pathology, ELISA detection of IL-6 levels, and RT-qPCR/Western blot analysis of IL-6, JAK2, and STAT3 mRNA and protein expression levels.

Results: The core targets of Dewetting and Pain-Relieving Powder in treating acute gastroenteritis (GA) were JAK2, STAT3, and IL-6, with active components primarily including paeoniflorin, mulberry polyphenols, quercetin, hyperoside, and hexahydrocurcumin. The involved signaling pathways included the JAK-STAT pathway and HIF-1 pathway, which are closely related to biological processes such as inflammatory responses and apoptosis. Molecular docking results demonstrated that the core targets (JAK2, STAT3, IL-6) and the active component paeoniflorin, which had the highest degree value in the network graph, exhibited favorable binding affinity. Animal experiments revealed that at 2 hours post-modeling, the ankle joint swelling rate increased in all groups and peaked at 12 hours before decreasing; the thermal pain threshold decreased, reached its lowest point at 2 hours, and then increased. At 72 hours post-modeling, compared to the model group, the colchicine group and high-dose CSDT group showed significant improvement in swelling severity and thermal pain threshold (P<0.05). Inflammatory responses in ankle synovial cells were alleviated across all colchicine and CSDT dose groups. Serum uric acid levels and IL-6 levels, as well as IL-6, JAK2, and STAT3 mRNA and protein expressions in synovial tissues, were significantly reduced in the colchicine group and high-dose CSDT group (P<0.05).

Conclusion: CSDT exerts therapeutic effects by regulating IL-6 signaling pathways through multi-component and multi-target mechanisms, thereby mitigating inflammatory responses and improving synovial tissue damage and disease progression in acute GA mouse models.