Jianpi Bushen Qingchang Huashi formula alleviates DSS-induced colitis through butyrate-associated HDAC1 inhibition and Th17/Treg homeostasis.

ETHNOPHARMACOLOGICAL RELEVANCE: Jianpi Bushen Qingchang Huashi Formula (JBQHF) is a traditional Chinese medicine compound of nine herbal ingredients clinically used to treat chronic diarrhea and dysentery syndromes, corresponding to ulcerative colitis (UC). Although clinical evidence supports its efficacy, the underlying mechanism remains unclear. AIM OF THE STUDY: This study sought to define the therapeutic benefit of JBQHF in a murine colitis model and to dissect the underlying mode of action. MATERIALS AND METHODS: Ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) was applied to profile the chemical constituents of JBQHF. Therapeutic efficacy was evaluated in a DSS-induced murine colitis model. Gut bacterial composition and metabolite output were analyzed by 16S rRNA gene sequencing and Gas chromatography-mass spectrometry (GC-MS), respectively. Network pharmacology, deep learning-based virtual screening, and molecular docking were employed to predict candidate active compounds and their potential targets. Immunological assays focused on Histone deacetylase 1 (HDAC1) expression, HDAC enzymatic activity, histone acetylation status and the T helper 17 / regulatory T (Th17/Treg) differentiation axis. RESULTS: JBQHF attenuated colitis as reflected by lower disease activity indices (DAI) and reduced histopathologic injury. Pro-inflammatory mediators were decreased while intestinal tight junction protein levels were elevated. JBQHF was associated with increased gut microbiota-derived butyrate production, reduced colonic HDAC1 expression and HDAC enzymatic activity, elevated Acetyl-H3K9 and Acetyl-H3K27, and a rebalancing of the Th17/Treg ratio. The protective effects of JBQHF were largely abrogated by the HDAC activator ITSA-1. Deep learning-based virtual screening further identified flavonoids within JBQHF as candidate HDAC1 ligands with predicted target selectivity. CONCLUSIONS: JBQHF ameliorates experimental colitis via a dual inhibition of HDAC1, providing an integrated mechanism for its clinical application.