Abstract: Objective: To investigate anti-inflammatory and analgetic effect of the ergosterol from fungi on nitroglycerin (NTG)-induced migraine in rats. Methods: Oral administration of ergosterol (15 and 30 mg/kg), rats were injected with NTG to establish migraine animal model, after nitroglycerin induction, their typical symptoms were observed and the levels of NO, interleukin-6 (IL-6), interleukin-1β (IL-1β), 5-hydroxytryptamine (5-HT), norepinephrine (NE) and dopamine (DA) in serum and brain tissue were examined by ELISA. The alteration of NF-κBp65, JAK2 and STAT3 in brain were determined by Western blot (WB). Results: After injected with nitroglycerin, the rats scratched their heads frequently. Compared with the model group, the levels of NO, IL-6 and IL-1β were significantly reduced in serum by different doses of ergosterol, and 5-HT, NE and DA in brain tissue were increased. WB results showed that the expressions of P-NF-κBp65, P-JAK2 and P-STAT3 were significantly decreased by different doses of ergosterol (P < 0.001). Conclusion: Ergosterol has a significant anti-inflammatory and analgetic effect on migraine. The mechanisms may be connected with NF-κB and JAK/STAT signaling pathways.