Key targets of brain-gut coordination in Wuzhuyu decoction treating chronic migraine based on proteomics

Chronic migraine is a debilitating disorder often accompanied by gastrointestinal dysfunction. However, the targets and mechanisms underlying this interaction in migraine remain unclear. Wuzhuyu Decoction (WZYD), a traditional Chinese medicine, has shown efficacy in alleviating migraine and gastrointestinal symptoms. However, its mechanisms of brain-gut coordination remain unclear. This study investigates the mechanisms of WZYD on the brainstem and colon in chronic migraine model rats based on proteomics. A chronic migraine model was induced via meningeal inflammation, followed by WZYD administration. WZYD increased pain thresholds, reduced CGRP and Fos, while elevating 5-HT. Proteomic analysis identified proteins significantly regulated by WZYD in the brainstem and colon respectively, primarily involved in inflammation and signal transduction. UHPLC-Q-Exactive-MS/MS revealed blood-absorbed components of WZYD, and network analysis predicted 959 potential targets. Proteomics-based PPI analysis combined with network analysis of blood-absorbed components identified DDO, PRKACB, and MAPK8 as key potential targets regulated by WZYD. These findings were validated through molecular docking, mRNA and protein expression analysis. WZYD reduced Iba-1, TNF-α, IL-6, SP, DDO, PRKACB, and MAPK8, while elevating IL-10 and GLP-2 levels, mitigating neuroinflammation and intestinal inflammation. These findings suggest WZYD could alleviate chronic migraine by regulating key protein targets and coordinating anti-inflammatory responses in the brain and gut.