Introduction & objective: Methotrexate (MTX) is a chemotherapeutic agent that used for the treatment of a variety of tumors and inflammatory diseases. Methotrexate cause defective oogenesis and spermatogenesis, and impair fertility. The aim of this study was planed to see the role of human growth hormone (GH) on testis recovery after treatment with methotrexate in rat. Methods The forty male wistar rat were selected and randomly divided into four groups; control (n=10) and test (n=30). GH groupe was received 0.3mg/kg GH (IP) for 28 consequence day, MTX Group was received 1mg/kg MTX (IP) per week for four consequence week, GH/MTX group was received 0.3mg/kg GH (IP) for 28 consequence day plus 1mg/kg MTX (IP) per week for four consequence week, however the control group just received normal saline (IP). In 14 and 28 days five rats from each group were sacrificed and the testes of rats in whole groups were removed and sperm was collected from epididymis then prepared for analysis. Results: Methodtrexate with 1mg/kg/week dose can cause histopatological and morphological changes in testicular tissue, damage of germinal cells and atrophy of seminiferous tubules. Also cause significant increase of interstitial tissue and capsular thickness and there was significant decrease (p<0.05) in testicular and body weight between this group and others specially control group. Administration of methotrexate cause significant decline in seminiferous tubules diameter and seminiferous epithelium thickness (p<0.05). There were no obvious changes seen in morphmetrical parameters between MTX/GH and control groups. In group that was treated with methodtrexate results shows significant decrease (p<0.05) in number, motility and viability of spermatozoa and testosterone concentration. Administration of GH in addition with methotrexate reduced the effects of methotrexate on sperm parameters and testosterone concentration even completely or partially. Conclusion These results suggested that GH had protective effect on approximately all destructive effects caused by MTX in rat testis and improved sperm parameters.