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HIF-VEGF pathway in hypoxia condition in middle ear due to dysfunction of eustachian tube is still unknown, which is considered as one of pathogenesis in otitis media.This study was designed to investigate the possible involvement of HIF-VEFG pathway in dysfunction of eustachian tube-induced otitis media with effusion.We adopted a soft palate approach to obstruct the orifice of eustachian tube to establish otitis media rat model.ABR, tympanometry were used for hearing function test, hypoxia related factors were examined by RT-PCR.The expressions of the hypoxia related proteins were detected by western blot and immunostaining.The model of otitis media with effusion was successfully induced by cauterizing the orifice of eustachian tube.RT-PCR showed upregulation of hypoxia related factors in cauterization-treated ears.Western blot and immunostaining showed that the expression of hypoxia related proteins in cauterization-treated ears were increased.Hypoxia-induced vascular proliferation and increasing of permeability may be one of pathogenesis of otitis media due to dysfunction of eustachian tube.