Essential oils from <i>Melissa officinalis</i> and <i>Citrus reticulata</i> ameliorate testicular inflammation and apoptosis induced by sleep deprivation

Sleep deprivation is a major risk factor for male infertility, largely via inflammatory and apoptotic pathways, yet effective treatments are limited. This study examined the protective effects of bioactive essential oils from Melissa officinalis and Citrus reticulata against testicular injury induced by total sleep deprivation (TSD). Forty-eight male Sprague Dawley rats were randomly assigned to six groups and exposed to TSD for 20 days using the multiple platform method. Interventions included daily inhalation of individual oils, their combination, or Alprazolam (ALP) as a control. Histopathological and immunohistochemical analyses (Cas-3, CD11b, NF-kappa B) were performed on testes, epididymis, and prostate. TSD caused severe structural degeneration in reproductive organs, with marked upregulation of apoptotic and inflammatory markers. Both essential oils alleviated these changes, while their combination provided superior protection compared to single use. Interestingly, the prostate showed relative resistance to TSD-induced injury compared with testes and epididymis. To our knowledge, this is the first evidence that inhalation of Melissa officinalis and Citrus reticulata essential oils significantly protects against sleep deprivation - induced reproductive toxicity. These findings suggest a safe, non-invasive aromatherapeutic strategy for preserving male fertility under chronic sleep stress.