OBJECTIVE: To study the short- and long-term effects of multi-glycosides of tripterygium wilfordii (GTWon) the histological structures of testes in pubertal rats and possible mechanisms. METHODS: Forty-eight 5-week-old male Sprague-Dawley rats were randomly intragastrically administered with low-does GTW（6 g/kg daily）and high-does GTW (12 mg/kg daily) or 1% sodium carboxymethylcellulose (6 mL/kg, control group) for four weeks. The testes were sampled for detecting histological structures and c-kit expression by immunohistochemistry 24 hrs and four weeks after drug discontinuance. RESULTS: The number of spermatogenic cells and the expression of c-kit in testes were reduced in the two GTW treatment groups 24 hrs and 4 weeks after drug discontinuance compared with those in the control group(P<0.05). Four weeks after drug discontinuance atrophy and interstitial edema of seminiferous epitheliumin in testes were observed, and the testis weight and the expression of c-kit in testes were reduced in the high-does GTW group compared with those in the control group (P<0.01). There were no significant differences in the parameters observed between the low-dose GTW and the control group 4 weeks after drug discontinuance. CONCLUSIONS: GTW has adverse effects on testes in a dose-dependent manner in puberty rats. Low-dose GTW may cause reversible short-term injuries to testis tissues. The damage of the interstitial tissue of testes induced by high-dose GTW may be one of the causes of long-term injuries of testes.