Xiongzhi Qufeng Zhitong Granule alleviates nitroglycerin-induced migraine-like nociception and central neuroinflammation by regulating the HMGB1 / TRPV1 / MAPK signaling axis.

ETHNOPHARMACOLOGICAL RELEVANCE: Chronic migraine (CM) represents a highly prevalent neuroinflammatory disorder with limited therapeutic options. Xiongzhi Qufeng Zhitong Granules (XZQF), a clinically used traditional Chinese medicine, displays promising anti-migraine potential with favorable safety profiles; however, its systematic efficacy and underlying mechanisms remain poorly defined.

AIM OF THE STUDY: This study aimed to investigate the protective effects of XZQF on a CM model in rats triggered by nitroglycerin (NTG) injection, and to unveil the potential mechanisms focusing on HMGB1/TRPV1/MAPK signaling pathway.

MATERIALS AND

METHODS

A CM rat model was first established, and the effects of XZQF on CM were evaluated integrating behavioral testing, enzyme-linked immunosorbent assay (ELISA), and histopathological staining analysis. Subsequently, anti-CM mechanism verification, including network pharmacological analysis, immunofluorescence, immunohistochemistry, and Western blotting, were employed to reveal the molecular mechanism of XZQF in regulating HMGB1/TRPV1/MAPK signaling axis to against CM.

RESULTS

Both behavioral assays, ELISA test, and histopathological assessment demonstrated that XZQF significantly restored body weight and pain thresholds, reduced serum and brain levels of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α), pain modulators (PGE2, 5-HT, β-EP), and vasoactive mediators (CGRP, VIP, NO, iNOS), while alleviating migraine-associated brain tissue damage. Network pharmacology identified core targets (STAT3, NFKB1, JUN, PTGS2, IL1B) and key bioactive components (ferulic acid, senkyunolides E/F, neocnidilide, methyleugenol), with enrichment in MAPK, PI3K-Akt, and cAMP signaling cascades. Immunofluorescence, immunohistochemistry, and Western blotting further validated that XZQF markedly suppressed the expression of CGRP, TRPV1, c-Fos, COX-2, and HMGB1, as well as the phosphorylation of MEK and MAPK.

CONCLUSION

Collectively, these findings demonstrate that XZQF exerts robust analgesic, anti-inflammatory, and vasoregulatory effects against CM by blunting central neuroinflammation through the HMGB1/TRPV1/MAPK signaling axis. Our work establishes a mechanistic framework for understanding the anti-CM activity of XZQF and supports its further development as a therapeutic intervention for CM.