Multi-omics analysis revealed that Jingfang Mixture alleviates inflammation in an OVA/Al(OH)3-induced urticaria-like rat model by regulating the AMPK/p38 MAPK/PI3K-Akt pathway associated with the gut-skin axis

Background The systemic and relapsing nature of urticaria cannot be fully explained by cutaneous mast-cell activation alone. Whether the gut contributes, and whether a multi-component formula such as Jingfang Mixture (JF) acts through a gut-skin route, have not been tested together at the level of microbiota, metabolites, and skin signaling together. Methods Using an OVA (ovalbumin)/Al(OH) 3 -induced urticaria-like allergic inflammation rat model, we integrated 16S rRNA gene sequencing, fecal untargeted metabolomics, and skin DIA (data-independent acquisition)-based quantitative proteomics to investigate the therapeutic mechanisms of JF. Results JF suppressed both Th2 (IgE (immunoglobulin E), IL-4 (interleukin-4)) and Th17/pro-inflammatory (IL-6 (interleukin-6), TNF-α (tumor necrosis factor-α), IL-17 (interleukin-17)) responses, improved intestinal barrier-associated abnormalities, and partially reversed gut dysbiosis by restoring SCFA-producing bacteria ( Bacteroidia, Clostridia, Lachnospirales, Oscillospirales ) while reducing Proteobacteria and Enterobacterales . JF normalized the fecal metabolome, elevating anti-inflammatory metabolites including butyrate, indole, indole-3-acetic acid, docosahexaenoic acid (DHA) and reducing pro-inflammatory mediators such as arachidonic acid, leukotriene B4, secondary bile acids. At the skin level, JF activated AMPK (AMP-activated protein kinase) signaling and suppressed aberrant p38 MAPK (p38 mitogen-activated protein kinase) and PI3K (phosphoinositide 3-kinase)-Akt (protein kinase B) activation. Spearman correlation analysis further revealed that SCFAs (short-chain fatty acids) and indole derivatives were positively correlated with the abundance of AMPK-pathway-related proteins and negatively with p38 MAPK and PI3K-Akt proteins, whereas pro-inflammatory metabolites exhibited inverse correlation patterns. Conclusion Collectively, our findings indicate that JF mitigates urticaria-like inflammation, likely through gut microbiota and modulating SCFA, bile acid, tryptophan, and lipid metabolism. This multi-omics evidence suggests that JF may act through the gut microbiota-metabolite-skin signaling axis to confer anti-urticarial benefits. Nevertheless, further causal validation experiments and clinical investigations are warranted.