Current data do not support the routine use of any agent to prevent cisplatin ototoxicity. Although there are various diseases in which derivatives of vitamin A are used due to their antioxidant effects, there is no study for prevention from ototoxicity. In this study, the protective effect of isotretinoin was investigated on cisplatin ototoxicity in rats. 21 Wistar Albino rats were divided randomly into 3 groups. Group I: cisplatin, Group II: cisplatin + isotretinoin and Group III was the control group. Hearing assessment of all rats was done with ABR and DPOAE tests before and after the procedure. After the procedure, cochleas were resected and transmission electron microscopic examination was performed. Our DPOAE and ABR findings showed that isotretinoin has protective effects on cisplatin ototoxicity. According to transmission electron microscopic findings, isotretinoin has protective effects on cell integrity. We think that new experimental and clinical studies to be carried out in this regard may give us a new option on prevention of cochlea from ototoxicity.