Background and Objective: Saffron is widely used as a food flavor and has well known medicinal effects. Recent studies have revealed that main components of saffron including carotenoids: crocin, crocetin, picrocrocin and safranal have a large number of physiological effects on different biological systems. We decided to assess the possible effect of saffron extract on concentratians of LH, FSH and testosterone in mice. Materials and Methods: Four groups each including eight adult male Balb/C mice weighing 30 5g were chosen. Normal saline was administered as placebo to control group and saffron extract in doses of 25 , 50 and 100 was injected intraperitoneally for 20 days to test groups. Serum FSH, LH and testosterone, were measured using ELISA. Results: The level of FSH, LH and testosterone significantly increased in 100 saffron treated group, as compared with with placebo group. No significant differences were observed between other test groups and placebo. Conclusion: The study indicates efficacy of saffron extract in dose of 100 mg/kg/ 48h on pituitary-testis axis in mice. However further studies are needed to determine the effect of saffron on human reproductivity.