Objective To investigate the target sites for male rats infertility by administered with combination regimen of lowdose gossypol acetic acid (GA) with steroid hormones (desogestrel/ethinylestradiol/testosterone undecanote (DSG/E/TU). Methods Adult male rats were randomly divided into four groups. Group GH: rats were fed orally with GA (125mg(kgI>&#/I>8226;day)) and DSG (0.125mg/(kgI>&#/I>8226;day))/E(0.025mg/(kgI>&#/I>8226;day))/TU (100mg/(kgI>&#/I>8226;day)); Group G: rats were administered with a single dose of GA (12.5mg/(kgI>&#/I>8226;day)); Group H: DSG(0.125mg/(kgI>&#/I>8226;day))/E(0.025mg/(kgI>&#/I>8226;day))/TU (100mg/(kgI>&#/I>8226;day)) were administered to rats; Group C: rats only received vehicle (1% methyl cellulose). The testis and epididymis were removed at 4, 6, 8, 10 weeks respectively. Morphology of testis, epididymis and sperm were observed, and testicular sperm number was counted, epididymal sperm motility was measured. Results We found that group GH had similar results as group H ,but not either as group G or as group C, in testicular sperm count and the morphology of testis seminiferous tubules, while in epididymis, the sperm motility decreased and the abnormal sperm increased obviously in all three treatment groups compared with group C.Conclusion The steroid hormones are the main functional factors on spermatogenesis in testis, gossypol and the hormones have synergetic effects on sperm motility in epididymis. With either independent or synergistic actions of gossypol and steroid hormones, the combined regimen have a promising effect on male rat contraception.