Spatial metabolomics combined with MALDI-MSI unveils gut-brain axis mechanisms of angelica dahurica radix in migraine rats.

BACKGROUND

Migraine is a highly disabling neurological disorder characterized by recurrent headaches and complex metabolic disturbances, affecting approximately 15% of the global population. Angelica dahurica Radix (Baizhi), a classic traditional Chinese medicine documented in Shennong Bencao Jing, has been used for millennia to treat migraine, but its underlying mechanism remains unclear.

PURPOSE: This study aimed to explore the gut-brain axis mechanisms underlying Baizhi's anti-migraine effects by integrating multi-omics approaches to clarify its regulatory effects on intestinal barrier function, gut microbiota, and cerebral metabolism.

STUDY

DESIGN

A nitroglycerin (NTG)-induced migraine rat model was used, with 48 male Sprague-Dawley rats randomized into four groups (n=12 each): Control (CON, 10 mL/kg water p.o.), Migraine model (NTG, water + NTG 10 mg/kg s.c.), Positive control (SS+NTG, sumatriptan succinate 260 mg/kg p.o. + NTG), and Baizhi aqueous extract (CBZ) group (7.2 g/kg crude drug + NTG). Migraine-like behaviors and biological samples were assessed after 22 days of intervention.

METHODS

The study combined MALDI mass spectrometry imaging (MALDI-MSI)-based spatial metabolomics (to analyze brain metabolic changes), fecal metabolomics (to identify systemic metabolic perturbations), and 16S rRNA sequencing (to profile gut microbiota composition). Additionally, histopathological staining (H&E) and immunohistochemistry (for Occludin and ZO-1) was used to evaluate intestinal barrier function, and ELISA(Enzyme-linked immunosorbent assay) was applied to detect pain-related biomarkers (5-HT, CGRP, TNF-α) in plasma and brain.

RESULTS

CBZ alleviated migraine-like behaviors (reduced head-scratching frequency) and normalized levels of pain-related biomarkers (5-HT, CGRP, TNF-α) in plasma and brain. It repaired intestinal barrier dysfunction by upregulating tight junction proteins (Occludin, ZO-1). 16S rRNA sequencing showed that CBZ reversed NTG-induced gut microbiota dysbiosis, specifically enriching Lactobacillus and reducing pro-inflammatory taxa (e.g., Blautia). Fecal metabolomics indicated restoration of key pathways, including lipid metabolism, glutathione metabolism, and short-chain fatty acid synthesis. MALDI-MSI visualized spatial metabolic normalization in the brain, with CBZ correcting abnormal glucose accumulation, restoring energy metabolism (e.g., ATP/GTP ratios), and regulating lipid signaling (e.g., phosphatidylinositol 4,5-bisphosphate) in specific brain regions.

CONCLUSION

Baizhi alleviates migraine through multi-target regulation of the gut-brain axis, including intestinal barrier repair, gut microbiota remodeling, and cerebral metabolic restoration. Notably, this study on Baizhi represents the first integration of spatial metabolomics (MALDI-MSI) with gut microbiome and systemic metabolomics to elucidate migraine mechanisms, providing a novel multi-omics strategy for understanding the holistic therapeutic effects of traditional Chinese medicine. These findings provide a mechanistic basis for its clinical application in migraine treatment.