Background Migraine is a common primary headache characterized by paroxysmal features and associated symptoms that significantly disrupt patients’ daily lives, work, and social interactions, rendering it the second leading disease cause of disability worldwide. Toutongping granule (TTP) is a well-established compound prescription in Traditional Chinese Medicine (TCM) for the treatment of migraine. However, the underlying mechanisms remain unclear. Objective This study aims to investigate the multi-pathway mechanisms through which TTP exerts its effects on migraine. Methods The therapeutic effects of TTP on migraine were assessed through behavioral observations, Nissl staining, Iba-1 staining and immunofluorescence (IF) for reactive oxygen species (ROS). Additionally, enzyme-linked immunosorbent assay (ELISA) was employed to measure serum cytokines (TNF-α, IL-6 and CGRP). The mechanism of TTP was examined in a nitroglycerin-induced migraine rat model using IF (CGRP, RAMP1, AKT, mTOR, p38 MAPK, p-STAT1, NRF2 and GPX4), Western blotting (PKA, cAMP, and p-CREB), and qRT-PCR ( miRNA382-5p, circNR3C1, TACR1, HSP60, and ATF4 mRNA) to evaluate neuroprotective, anti-inflammatory, and antioxidant effects. Results TTP reduces headache-like behaviors, diminishes mitigates neuronal damage, inhibits aberrant activation of microglia and ROS production, decreases pro-inflammatory cytokines (TNF-α, IL-6), and modulates key signaling proteins (CGRP, RAMP1, AKT, mTOR, p38 MAPK, p-STAT1, NRF2, GPX4, cAMP, PKA, p-CREB) along with genes ( miRNA382-5p, circNR3C1, TACR1, HSP60, and ATF4 mRNA) associated with migraine. Conclusion TTP demonstrates multi-pathway and targets, exhibiting neuroprotective, anti-inflammatory, and antioxidant effects, thereby supporting its application in migraine treatment.
Zhao et al. (Sun,) studied this question.