Synergistic Multi-Target Therapy: berberine-Taxifolin Co-Administration Attenuates Murine Inflammatory Bowel Disease via NF-κB/NLRP3 Suppression and Intestinal Barrier Restoration

Objective: To investigate the cooperative therapeutic effects of berberine and taxifolin, two anti-inflammatory phytochemicals, in a murine colitis model. Methods: Network pharmacology was used to explore the potential therapeutic effects and mechanisms of berberine and taxifolin combination in treating colitis. A colitis model was established in mice using 3% dextran sulfate sodium (DSS) administered via drinking water. Mice were treated with berberine alone, taxifolin alone, or a combination of both, with daily monitoring of body weight changes. Hematoxylin-eosin (HE) staining was performed to evaluate intestinal structural integrity and inflammatory cell infiltration. TUNEL assay and western blotting (WB) were utilized to detect apoptosis levels in intestinal cells, with quantification of Caspase3 p17/p19, Bax, and Bcl-2 protein expression. Immunofluorescence staining was conducted to assess the expression of tight junction proteins (occludin and ZO-1) and MUC2, reflecting intestinal barrier function. Pro-inflammatory cytokines (IL-1β, IL-6, TNF-α, and iNOS were measured by WB. Additionally, immunofluorescence staining of F4/80 ⁺ macrophages was performed to quantify inflammatory cell infiltration in intestinal tissues. Potential therapeutic targets of the berberine-taxifolin combination were further investigated through molecular docking to elucidate synergistic mechanisms. Results: Network pharmacology findings suggest that the combined use of berberine and taxifolin may modulate pathways associated with programmed cell death, immune response regulation, and apoptosis in colitis. Experimental results demonstrated that both berberine and taxifolin individually attenuated DSS-induced body weight loss and colorectal shortening, with their combination exhibiting synergistic inhibition. HE staining confirmed that the combination therapy more effectively restored intestinal structural integrity. Comprehensive analyses revealed that individual administration of berberine or taxifolin suppressed pro-apoptotic proteins (Caspase3 p17/p19 and Bax), upregulated anti-apoptotic Bcl-2 expression, and reduced TUNEL-positive cell counts, while their combination further amplified these anti-apoptotic effects. Additionally, the combination significantly enhanced the expression of ZO-1, occludin, and MUC2, indicating superior restoration of intestinal barrier integrity. Both compounds alone reduced intestinal levels of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) and iNOS, as well as decreased F4/80 ⁺ macrophage infiltration, with their combination synergistically suppressing inflammatory responses. Molecular docking simulations further identified NF-κB1, NLRP3, PPAR-γ, and STAT3 as potential direct binding targets of berberine and taxifolin, suggesting these interactions may underlie their combined therapeutic mechanisms. Conclusion: These findings established the combined use of berberine and taxifolin as a novel multi-target therapeutic strategy that concurrently addresses inflammatory dysregulation, barrier repair, and apoptosis control in inflammatory bowel disease (IBD), providing a phytochemical synergy blueprint for complex inflammatory disorders.