Xanthohumol alleviates central sensitization in migraine via PPARα-mediated activation of the AMPK pathway and suppression of neuroinflammation.

Abstract

BACKGROUND: Migraine is a common neurological disorder characterized by repeated attacks, with central sensitization recognized as a critical mechanism underlying the pathogenesis of chronic migraine (CM). Xanthohumol (Xn), a predominant bioactive allyl flavonoid derived from hops (Humulus lupulus L.), demonstrates notable anti-inflammatory and antioxidant properties. However, its therapeutic potential for migraine remains poorly characterized. PURPOSE: The study focused on investigating the protective effects of Xn on a migraine model in mice, which was triggered by repeated dural applications of inflammatory soup (IS), and aimed to identify the underlying mechanisms. METHODS: A migraine mouse model was established through repeated dural IS stimulation. These mice received two doses of Xn treatment. Headache behavior was assessed using tests including head scratching, the periorbital mechanical threshold, cold allodynia, and light aversive behavior. Analyses of biomarkers and key protein expression via Western blot (WB), immunofluorescence (IF), and immunohistochemistry (IHC) were conducted to evaluate central sensitization. Through network pharmacology, molecular docking, cellular thermal shift assay (CETSA), and surface plasmon resonance (SPR), peroxisome proliferator-activated receptor α (PPARα) was identified as a potential target for Xn's migraine treatment. Downstream molecular mechanisms were validated by both activating and inhibiting PPARα. RESULTS: Comprehensive behavioral analyses revealed that Xn administration significantly alleviated migraine-like symptoms in migraine model mice, including increasing the periorbital mechanical threshold, reducing cold allodynia and head scratching, and improving light aversive behaviors. Network pharmacological analyses suggested that PPARα was a potential target for Xn in the treatment of migraine. Molecular docking, CETSA, and SPR experiments confirmed direct binding between Xn and PPARα. Mechanistic investigations demonstrated that Xn exerted regulatory effects through PPARα/AMPK pathway activation coupled with cyclooxygenase-2 (COX-2) suppression and downregulation of pro-inflammatory cytokines. CONCLUSION: This study revealed that Xn alleviated migraine-related central censitization by activating the PPARα/AMPK axis and reducing neuroinflammation, proposing Xn as a novel migraine treatment.