Fluoxetine is a selective serotonin reuptake inhibitor (SSRI), it is used in the treatment of depression; it has a toxic effect on the testis. Major depressive disorder is a pathological disorder associated with increased levels of the inflammatory cytokines tumor necrosis factor-alpha (TNFα), interleukin-6 (IL-6), and IL-1 beta (IL-1β). Fluoxetine increase in malondialdehyde (MDA) and decrease in antioxidant enzyme activity SOD superoxide dismutase and catalase (SOD and CAT) and reduced glutathione (GSH).The study aimed to investigate whether vitamin D can reduce the oxidative damage caused by fluoxetine.  Thirty -two adult male rats are divided into four groups and are included in the analysis. Animals in Group I (control group) were administered distilled water by gavage. Groups II: animals were given a dose of 10 mg/kg of fluoxetine (fluoxetine treated group) orally by gavage daily for 4 weeks. Group III (vitamin D group) animals received intramuscular VD (1,000 IU/kg; 3 days/week for 4 weeks. Group IV (fluoxetine+ vitamin D): drugs were given in the same previous doses for 4weeks. Blood samples were obtained 24 hours after the last dose of each drug.The biochemical results showed that fluoxetine significantly increased oxidative stress in testicular tissue and inflammatory markers in serum. The in-depth investigations supported that administering the fluoxetine combined with vitamin D reduced the testicular damage to a marked level and normalized all relevant markers. It was concluded thatthe oxidative stress induced by fluoxetine administration in rats could be reduced by vitamin D supplementation.