Background <p>Medication overuse headache (MOH) is a chronic disorder due to excessive acute headache treatment use. Neuroimaging studies suggest the involvement of the orbitofrontal cortex (OFC) in the pathophysiology of MOH, particularly its connections with the periaqueductal gray (PAG). The OFC-vlPAG circuit has been well-established as a critical neural substrate in pain regulation; while its role in triptan-related MOH pathogenesis remains unclear. This study aims to investigate the role of the OFC-vlPAG circuit and its underlying mechanisms in regulating mechanical hypersensitivity using a triptan-induced MOH mouse model.</p> Methods <p>Male C57BL/6J mice (along with a female exploratory cohort) were used to develop a triptan-induced MOH model through repeated rizatriptan (RIZ) administration. Behavioral assessments of cutaneous allodynia were conducted using von Frey filaments. Immunofluorescence staining was performed to examine neuronal activity and 5-HT_1D receptor (5-HT_1DR) expression in the OFC. Chemogenetic and optogenetic techniques were employed to modulate vlOFC glutamatergic neurons or the vlOFC-vlPAG pathway, and pharmacological methods were utilized to target the vlOFC’s 5-HT_1DR.</p> Results <p>Repeated RIZ administration induced cutaneous allodynia in the MOH mouse model, with significant reductions in hind paw and head withdrawal thresholds. Elevated c-Fos expression in the vlOFC CaMKII-α⁺ neurons of triptan-induced MOH mice indicated increased glutamatergic neuronal activity. Chemogenetic and optogenetic glutamatergic neuron activation in the vlOFC alleviates allodynia in the triptan-induced MOH model. Furthermore, glutamatergic vlOFC-vlPAG circuit activation significantly improved pain thresholds in these MOH mice. The observed downregulation of 5-HT_1DR in the vlOFC of these MOH mice was functionally associated with inhibition relief in this circuit. This adaptation may create a permissive state, allowing for robust analgesic effects upon targeted exogenous activation.</p> Conclusions <p>Activating the glutamatergic OFC-vlPAG circuit elicits robust analgesia in triptan-induced MOH mice. 5-HT_1DR downregulation is hypothesized to unmask the circuit’s analgesic potential by elevating vlOFC baseline activity, providing a highly responsive therapeutic target for triptan-related allodynia.</p> Clinical trail number <p>Not applicable.</p> Graphical Abstract <p></p>