Thymoquinone has antioxidant properties. We hypothesized that thymoquinone may prevent or alleviate hearing loss induced by acoustic trauma. We aimed to study thymoquinone's effect on hearing function with distortion-product otoacoustic emissions and auditory brainstem response. Thirty adult Spraque Dawley rats were randomized into four groups following exposure to acoustic trauma for 4 h. Control group (n = 7) did not receive further treatment. Thymoquinone-20 (n = 8) and Thymoquinone-40 (n = 8) received 20 and 40 mg/kg of intraperitoneal thymoquinone, respectively. Corn-oil group (n = 7) received 1 ml of corn oil intraperitoneally. Hearing function of both ears was tested with distortion-product otoacoustic emission and auditory brainstem response before, and shortly after acoustic trauma, and 96 h following acoustic trauma. Post-trauma signal/noise ratios and wave V amplitude/latencies of all groups were significantly low compared with pre-trauma values, which indicate no preventive effect of thymoquinone. Rats in Thymoquinone-20 showed a significantly improved distortion-product otoacoustic emission and auditory brainstem response results at 4000 frequency and above in post-treatment tests (p < 0.05). Improvement in Thymoquinone-40 at the same frequencies was insignificantly inferior to Thymoquinone-20, yet superior to control and corn-oil groups (p < 0.05). We conclude that thymoquinone may not prevent acoustic trauma-induced hearing loss, however, at 20 mg/kg for 96 h, may repair the damage.