Objective To study the effects of simulated weightlessness induced by tail-suspension on reproduction in adult male rats and its mechanisms. Methods Twenty Spraque-Dawley adult male rats were randomly divided into two groups: the experimental group and control group. Rats in the experimental group were tail-suspended for 14 days. After the treatment, the weight and morphology of testis, the quantity and quality of sperm, the level of serum testosterone and the apoptosis rate of testicular cells were observed. Results Compared with the control, the testicular weight, the quantity and quality of sperm, the level of serum testosterone significantly decreased (P＜0.05) while the apoptosis rate of testicular cells significantly increased (P＜0.05) in tail-suspended rats. In addition, hematoxylin and eosin (HE) staining showed that seminiferous tubules atrophied, spermatogenic cells decreased, seminiferous epithelial cells disintegrated and shed, and the lumina were azoospermic. Conclusion Simulated weightlessness induced by tail-suspension has a harmful effect on male reproduction of adult rats by increasing the apoptosis of testicular cells.