Elucidating the molecular mechanisms of Daifu decoction in ulcerative colitis treatment through a multi-omics framework and experimental verification.

Abstract

BACKGROUND: Ulcerative colitis (UC) is a significant and challenging condition in the digestive system, necessitating the development of effective therapeutic interventions. The Daifu decoction (DFD) is derived from the Lizhong decoction, a classic traditional Chinese Medicine (TCM) formula used to treat digestive diseases. Previous studies have found that DFD has a clear therapeutic effect on UC. This study aimed to investigate the underlying mechanisms of DFD in UC treatment. METHODS: A mouse model of dextran sulfate sodium (DSS)-induced UC was established, and mice were treated with DFD using the concentrations 1.6 g/(kg·d), 3.2 g/(kg·d), and 6.4 g/(kg·d) for 7 days. H&E and AB-PAS staining, ELISA, qPCR, and immunofluorescence were selected to assess the role of DFD in alleviating colitis and improving intestinal barrier damage. In addition, untargeted metabolomics and transcriptomics helped investigate the potential mechanism of DFD against UC, 16S rRNA sequencing evaluated the characteristics of the gut microbiota. The key target and downstream molecules were verified by molecular docking, molecular dynamics simulation, qPCR, western blotting, and immunohistochemistry. RESULTS: DFD prevented goblet cell loss, downregulated proinflammatory factors in serum, and improved tight junction mRNA and protein expression in colon tissue. Furthermore, DFD decreased the proportion of pathogenic microbes, while increasing the microbiota diversity and the proportion of beneficial bacterial. The integrated multi-omics analysis revealed that PRKCG was the key target of DFD to improve UC. Molecular docking and molecular dynamics simulations further confirmed that the active components of DFD could bind to PRKCG stably. The results of qPCR, western blotting, and immunohistochemistry revealed that DFD could regulate PKCγ/ERK/NF-κB pathway. CONCLUSION: DFD can exert therapeutic effects on UC by regulating the PKCγ/ERK/NF-κB signaling pathway and the gut microbiota to restore the intestinal barrier and decrease the secretion of proinflammatory cytokines.