Ligusticum sinense Oliv. cv. Chaxiong Extract Alleviates Nitroglycerin-Induced Migraine-like Responses in Rats: Associations with Glycerophospholipid Metabolism and PI3K/AKT Signaling

Ligusticum sinense Oliv. cv. Chaxiong (Chaxiong) has traditionally been used as a medicinal tea for headache relief in southern China, but its bioactive basis and mechanisms remain unclear. This study integrated ultra-performance liquid chromatography–quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS), untargeted metabolomics, network pharmacology, molecular docking, reverse-transcription quantitative PCR (RT-qPCR), and Western blotting in a nitroglycerin-induced migraine-like rat model. Chaxiong extract alleviated migraine-like behavior, reduced calcitonin gene-related peptide (CGRP), nitric oxide (NO), and tumor necrosis factor-α (TNF-α), increased 5-hydroxytryptamine (5-HT), and ameliorated brain histopathological alterations. A total of 48 constituents and 13 differential serum metabolites were annotated or putatively annotated, respectively. Glycerophospholipid metabolism was the principal metabolic pathway affected by treatment, whereas network pharmacology prioritized phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling. Pathway integration identified PIK3CA, PIK3CB, AKT1, AKT2, and INS as candidate bridging targets. Exploratory docking suggested potential interactions between five representative constituents and PIK3CA/AKT1. RT-qPCR showed reduced Pik3ca and Akt1 mRNA expression, while Western blotting showed decreased p-PI3K p85α (Tyr607)/PI3K p85α and p-AKT (Ser473)/AKT1 ratios following treatment. These findings suggest that Chaxiong-associated alleviation of migraine-like responses was accompanied by altered glycerophospholipid metabolism and reduced PI3K/AKT pathway activation.