Supporting data for “Effects of Angong Niuhuang Pill on ischemic stroke via the regulation of gut microbiota targeting TMA/TMAO metabolic pathway”

BackgroundIschemic stroke is a cerebrovascular event with underlying pathology in which gut microbiota and their metabolites participate via the gut-brain axis. The traditional Chinese medicine Angong Niuhuang Pill (AGNHP) is widely used in treatment of stroke, but its mechanisms via gut microbiota modulation remain obscure. ResultsIn this study, we established a middle cerebral artery occlusion (MCAO) mouse model and administered AGNHP at different timepoints after stroke. Multi-omics approaches including 16S rRNA sequencing and LC-MS/MS-based targeted metabolomics were conducted to analyze gut microbiota composition and TMA/TMAO metabolic pathways dynamically. The results showed that AGNHP treatment significantly reduced infarct volume, improved neurological function, relieve inflammatory conditions and restored gut and blood-brain barrier (BBB) integrity. Mechanistically, AGNHP altered gut microbiota composition. Through integrating abundance profiling, Lefse analysis, and correlation analysis related to metabolites, further study revealed AGNHP exclusively enriched the probiotic Akkermansia muciniphila, which suppressed TMAproducing-related genes (CntA/B, CutC/D, YeaW/X), thereby reducing systemic and cerebral TMA/TMAO accumulation. Complementary in vitro studies demonstrated that TMA may directly disrupted tight junction proteins in BBB and intestinal barrier. ConclusionsThis study elucidates the neuroprotective mechanism of AGNHP on ischemic stroke through modulation of the A. muciniphila-TMA/TMAO. By establishing a novel “Complex Drugs-Core Strains-Key Metabolites” paradigm, our study elucidates how AGNHP protects against ischemic stroke through time-dependent regulation of the gut microbiota-TMA/TMAO axis, with A. muciniphila playing a pivotal role. These findings not only advance our understanding of stroke pathophysiology, but also provide a methodological framework for investigating complex traditional medicines in modern biomedical research.