BACKGROUND: Glucocorticoid (GC) excess adversely affects male fertility. AIMS: This study examined the protective effects of Rosa damascena essential oil (RD oil) on testicular structure and function as well as sperm parameters in a rat model of GC-induced testicular injury. METHODS: Forty-nine adult rats were divided into seven groups: negative control (NC): normal rats; positive control (PC): received dexamethasone at 7 mg/kg/day (intraperitoneally for 7 days); vehicle control (VC) and RD oil-treated groups (T1-T4): received olive oil as vehicle or treated with 40, 80, 160 and 320 mg/kg/day of RD oil by oral gavages concurrently with dexamethasone. RESULTS: Dexamethasone decreased body weight and serum testosterone of PC rats, accompanied by germ cell depletion, epithelial thinning, and a marked increase in apoptotic cells in the testes compared to NC rats (P<0.05). In PC rats, reductions were observed in epididymal total sperm count, the number of morphologically normal and viable spermatozoa, sperm DNA integrity, and the percentage of acrosome-intact spermatozoa. In contrast, sperm kinematic parameters remained largely unchanged. RD oil, particularly at higher doses, positively affected testosterone levels, spermatogenesis, and seminiferous epithelial thickness, and reduced testicular apoptosis. In addition, RD oil improved certain sperm parameters, including the percentage of acrosome-intact sperms and DNA integrity. CONCLUSION: RD oil shows a protective effect against GC-induced injury to spermatogenesis and testicular tissue, as well as derangement in serum testosterone. These effects are associated with anti-apoptotic effects, especially at higher doses. Certain impaired sperm parameters are also improved by RD oil. These findings set the stage for future investigations on RD oil in GC-related infertility.