Cisplatin (CP) is one of important tumour chemotherapeutic agents in humans. Previous reports claim that CP can cause testicular toxicity. The aim of this study was to evaluate the potential effects of CP in the testes of rats. Male Wistar rats were intraperitoneally administered CP at 1.0, 2.5, and 5.0 mg/kg for three consecutive days. After exposure, CP significantly inhibited the testicular activities of succinate dehydrogenase (SDH) and malate dehydrogenase (MDH), but it significantly elevated the activities of acid phosphatase (ACP), alkaline phosphatase (AKP), and lactate dehydrogenase (LDH) in the 5.0 mg/kg group. The decreased levels of superoxide dismutase (SOD), total antioxidant capacity (T-AOC), and metallothionein-1 (MT-1) mRNA as well as the increased levels of malondialdehyde (MDA) and haemoxygenase-1 (HO-1) mRNA showed that CP could increase oxidative stress in rat testes. Western blot analysis showed that the levels of transferrin, vimentin, androgen binding protein (ABP) and inhibinβ-B decreased significantly in the CP 2.5 and 5.0 mg/kg groups compared with the control group. These findings indicated that the inhibited enzymes, oxidative stress, and the down-regulation of Sertoli cell function-related proteins play pivotal roles in CP-induced testicular damage.