Pinellia ternata–Coptidis Rhizoma herb pair suppresses Helicobacter pylori colonization via NLRP3-mediated pyroptosis

A B S T R A C T Ethnopharmacological relevance Pinellia ternata (Ban-xia) and Coptidis Rhizoma (Huang-lian) form a classic herb pair (BXHL) in traditional Chinese medicine that has demonstrated clinical efficacy against Helicobacter pylori ( H. pylori ) infection. However, its pharmacological mechanisms remain insufficiently defined. Aim of the study This study aimed to (1) evaluate the anti- H. pylori colonization effects of BXHL in vitro and in vivo; (2) elucidate its mechanism through the NLRP3-mediated pyroptosis pathway; and (3) assess its influence on gastrointestinal microbiota and identify potential active components. Materials and methods BXHL composition was characterized by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS). Anti- H. pylori activity was examined using GES-1 cell co-culture and infected mouse models. Gene expression and signaling pathways were analyzed by RNA-seq, ELISA, RT-qPCR, Western blotting and IHC. Histopathology, bacterial load assays, and ultrastructural observation assessed anti-colonization efficacy. Microbiota alterations were analyzed via 16S rRNA sequencing and molecular docking evaluated interactions between BXHL compounds and NLRP3 targets. Results BXHL significantly reduced H. pylori adhesion, colonization and restored NLRP3-mediated pyroptosis suppressed by infection. Transcriptomic and molecular analyses confirmed reactivation of the NLRP3 signaling cascade. BXHL preserved gut microbial diversity, unlike triple therapy, and exhibited moderate direct antibacterial effects. Conclusion BXHL inhibits H. pylori colonization through suppressing bacterial adhesion factors and activating NLRP3-mediated pyroptosis, while causing no disruption to the gut microbiota. These findings position BXHL as a safe and effective adjunctive therapy, which provides a novel strategy to improve H. pylori eradication that complements conventional antibiotics.