Noise-induced hearing loss (NIHL) significantly impacts the quality of life for patients. Extracellular vesicles (EVs) play crucial roles in cellular communication, but it is unclear if the inner ear releases EVs into peripheral circulation under NIHL conditions as well as specific constituents. The NIHL mouse model was established by exposing mice to continuous 120 dB SPL white noise for 2 h, and hearing loss was assessed with auditory brainstem response (ABR). EVs were isolated by size exclusion chromatography (SEC) and characterized. Protein profiling in plasma EVs from NIHL mice was assessed via mass spectrometry and validated by ELISA and Western blot. Inner ear Hsp70 expression was interfered with intra-tympanic delivery of siRNA-Hsp70 into the middle ear. Hsp70 expression in plasma EVs was determined by ELISA. Intraperitoneal N-acetylcysteine (NAC) was performed to understand the relationship between Hsp70 expression in plasma EVs and inner ear oxidative stress. Single-cell analysis identified potential sources of EV-associated Hsp70 in the inner ear. Stria vascularis endothelial cells (SV-ECs) were stimulated with HO, and EV-associated Hsp70 in supernatants was determined by ELISA. The NIHL mouse model was successfully established. Plasma EVs, isolated via SEC, exhibited membrane-bound nanoparticles, with CD63, Alix, and TSG101 markers identified. Proteomic analysis revealed increased Hsp70 expression in EVs from NIHL mice, which were further confirmed with ELISA and Western blot. Intra-tympanic delivery of siRNA-Hsp70 decreased expression of Hsp70 in plasma EVs, indicating the inner ear as one of the potential sources of plasma EV-associated Hsp70. NAC therapy reduced inner ear Hsp70 expression, suggesting its association with noise-induced oxidative stress. SV-ECs were identified as one of the key cells for secretion of Hsp70-carrying EVs. Hsp70 expression was significantly increased in plasma EVs from NIHL mice upon noise-induced inner ear oxidative stress, with SV-ECs potentially being key release sites, suggesting diagnostic and therapeutic applications for plasma EV-associated Hsp70 in NIHL. KEY MESSAGES: Hsp70 was significantly increased in plasma EVs of NIHL, which was partially derived from SV-ECs upon noise-induced oxidative stress. Plasma EV-Hsp70 could be the potential target for diagnosis and therapy of NIHL.