Background: Broccoli with scientific name, Brassica Oleracea, from Crucifera genus is a vegetable full of vitamins (such as A, C and E), fibers, Beta-carotene, calcium, iron, zinc, selenium and Flavonoids, which is a strong antioxidant and it can prevent from damages of Reactive Oxygen Species in body The present study investigated the effects of Brassica Oleracea extract on LH, FSH and Testestrone hormones levels.Methods: Forty adult male wistar rats were used. They were feeded from standard Pellets. The animals were divided into five groups: Control, Blank, Experimental 1, Experimental 2, and Experimental 3 (n=8). The plant extract doses used for experimental groups 1, 2, and 3 were 0.5gr/kg, 1gr/kg, and 2gr/kg respectively. Using gavage the hydro-alcoholic extract was orally administered to animals. After 28 days of feeding, the animals were made unconscious by ether, and blood samples were obtained from the heart. Then all groups’ blood serums were separated, and concentrations of LH, FSH and testosterone hormones were evaluated using Elisa Method.Results: In all groups, the levels of LH hormone had not significant increase, but the increased amount of Group 2 was more than the others; the levels of FSH had a significant decrease in the Experimental Group 3 as compared to Control Group. Concentration of testosterone in the Experimental Group 2 showed a significant increase as compared to Control Group.Conclusion: Findings showed that the effect of Brassica Oleracea extract was mostly presumed due to existence of flavonoids which results in increasing and decreasing of hormones, Flavonoids have a phytoestrogen structure that can be used in steroid synthetic cycle [in the other hand, they can be treated as a antioxidant which could eliminate ROSs in body].The investigation revealed that Brassica Oleracea hydroalcoholic extract has a dose-dependent activity. The obtained suitable dose in the study was 1gr/kg.