The present study explored the role of endogenous nitric oxide (NO) in the temporary threshold shift caused by acoustic trauma. Guinea pigs were exposed to broadband white noise at a level of 105+/-2dB sound pressure level (SPL) for 10min, causing a temporary threshold shift (TTS). The guinea pigs were divided into six groups (N-1 to N-6) according to survival days after noise exposure (0, 1, 2, 3, 7, 28days). Auditory brainstem responses (ABR) were recorded before noise exposure, immediately after noise exposure and before sacrifice. Immediately after animals were sacrificed, the stria vascularis and the spiral ligament of the lateral wall of each individual cochlea were harvest as a unit and prepared for assay of NO. There was a significant correlation (P<0.001) between the NO concentration and final ABR threshold in the noise exposure groups. But the return of ABR threshold to pre-noise-exposed level is early than that of NO concentration. An average 16.2dB threshold shift was found immediately after noise exposure. The threshold returned to the pre-noise-exposed level on the second post-exposure day. Comparing to unexposed control animals, the NO concentration increased nearly threefold immediately following noise exposure and decreased to twofold when the hearing threshold had returned to the pre-noise-exposed level. On the seventh post-exposure day the NO concentration was not different from that in unexposed control animals. Those findings indicate that endogenous NO is generated in the noise-induced temporal threshold shift and its concentration is correlated with the hearing loss.