An Herbal Enema Formula Alleviates Ulcerative Colitis by Inhibiting Enteric Glial Activation <em>via</em> the S100β/RAGE/NF-κB Pathway

Activation of enteric glial cells (EGCs) contributes to the pathogenesis of ulcerative colitis (UC). This study examined whether Dibai Enema Liquid (DBE)-an herbal enema combining Sanguisorba officinalis, Bletilla striata, and Acacia catechu-modulates EGC-driven inflammation. LC-MS/MS profiles of DBE constituents and prioritized compounds were docked to S100β. Efficacy was assessed in dextran sulfate sodium (DSS)-induced murine colitis using body weight, colon length, histopathology, and cytokine levels (ELISA). Glial activation markers were measured by immunohistochemistry, immunofluorescence colocalization, quantitative PCR (qPCR), and immunoblotting. An in vitro model used CRL-2690 enteric glia activated with lipopolysaccharide plus interferon-γ, with DBE-containing rat serum (15%) as the intervention. Docking suggested a strong predicted binding of several DBE components to S100β. DSS increased TNF-α and IL-1β, disrupted tight junctions, and elevated S100β/GFAP at both mRNA and protein levels, consistent with glial hyperactivation; DBE significantly reversed these changes. In vitro, DBE reduced S100β, RAGE, iNOS (NOS2), and phosphorylated NF-κB p65. Collectively, DBE appears to alleviate UC by inhibiting EGC activation and downregulating the S100β/RAGE/NF-κB/iNOS axis, thereby preserving mucosal integrity.