Potential Mechanism of Qiju Dihuang Pill in Treating Primary Sjogren's Syndrome Revealed by Network Pharmacology and Experimental Verification

Primary Sjogren's syndrome (pSS) presents as a persistent inflammatory condition marked by a spectrum of symptoms and a limited array of conventional therapeutic interventions. Within Traditional Chinese Medicine (TCM), the Qiju Dihuang Pill (QJDHW) stands as a frequently employed prescription for addressing this syndrome, yet the underlying therapeutic mechanisms remain elusive. Traditional Chinese Medicine Systems Pharmacology (TCMSP) and three disease gene databases, DisGnet, GeneCards, and OMIM, were utilized to establish QJDHW targets and pSS-related gene sets. Cytoscape was used to construct Protein-protein interaction (PPI) and active compound-target networks, followed by Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment via the DAVID database. Molecular docking was conducted with AutoDock Vina, and in vitro experiments verified the key findings. A total of 62 overlapping targets were identified. Network analysis revealed quercetin and kaempferol as major active compounds. KEGG enrichment indicated that QJDHW may exert its therapeutic effects by modulating the immune-inflammatory response. Four key targets, namely NFKBIA, IL-6, JUN, and IL-1B, were identified as the central mediators. Molecular docking analysis confirmed their binding affinity with the major active compounds. In vitro assays further confirmed that quercetin and kaempferol suppressed the mRNA expression of key inflammation-related factors, including IL-6, IL-1B, JUN, and NFKB, while reducing IL-6 and IL-1B protein levels and significantly inhibiting the phosphorylation of JUN and NFKB. Collectively, the integration of network pharmacology and experimental validation demonstrated that QJDHW exerts anti-inflammatory effects in treating pSS.