TRPC5 involvement in NTG-induced migraine-like central sensitization in mice: a study based on the PI3K-Akt/P2X7R-IL-1β signaling axis

Background Central sensitization is the core pathological mechanism driving the occurrence and chronification of chronic migraine (CM). Transient Receptor Potential Canonical 5 (TRPC5) channel is critically involved in the modulation of multiple pain disorders and central sensitization. However, the specific regulatory role and precise molecular mechanism of TRPC5 in the progression of acute and chronic migraine remain unclear. This study aimed to explore the potential function of TRPC5 and its downstream signaling cascade in nitroglycerin (NTG)-induced migraine-like central sensitization, so as to provide novel preclinical evidence for migraine pathogenesis and targeted intervention. Methods A repetitive NTG intraperitoneal injection protocol was applied to establish a mouse model mimicking acute migraine nociceptive responses and chronic migraine central sensitization. Migraine-like behavioral phenotypes, including mechanical allodynia, thermal hyperalgesia and light-aversive behavior, were evaluated at both acute and chronic modeling stages. qRT-PCR and Western blot were performed to detect TRPC5 mRNA and protein expression in the primary somatosensory (SSP) cortex. Immunofluorescence staining was used to verify the neuronal localization of upregulated TRPC5 and assess the activation status of cortical neurons and microglia. Pharmacological interventions with TRPC5 selective antagonist AC1903 and phosphoinositide 3-kinase-protein kinase B (PI3K-Akt) inhibitor LY294002 were applied to validate the regulatory relationship among TRPC5, PI3K-Akt signaling and purinergic receptor P2X7/interleukin-1 beta (P2X7R/IL-1β)inflammatory axis. Results Repeated NTG challenge induced robust migraine-like behaviors, prominent neuronal hyperactivation and sustained central sensitization within the SSP cortex. TRPC5 mRNA and protein were markedly increased mainly in SSP cortical neurons. NTG also triggered microglial overactivation and upregulated P2X7R and IL-1β. AC1903-mediated TRPC5 inhibition alleviated acute and chronic migraine-like phenotypes, blunted neuronal and microglial activation, suppressed PI3K-Akt overactivation and reduced P2X7R/IL-1β expression. LY294002 blockade of PI3K-Akt recapitulated these protective effects, attenuating sensory hypersensitivity and neuroinflammation. Conclusion TRPC5 mediates NTG-induced migraine central sensitization. Neuronal TRPC5 upregulation activates PI3K-Akt signaling, inducing neuronal hyperexcitability, microglial overactivation and P2X7R/IL-1β neuroinflammation. This TRPC5-dependent axis controls acute nociception and chronic migraine progression. Our findings uncover neuron-microglia crosstalk and support TRPC5 as a preclinical therapeutic target for migraine.