Objective To investigate the significance of p53/Rb cellular transduction pathway related genes and proteins in testis aging by detecting expressions of p53, Rb, p16, p19 and p21 in aging rat testis. Methods Thirty rats of 8-week-old male SD rats (weight: 180-220g) were randomly divided into 2 groups, 15 rats in each: the normal control group and the aging model group. The sub-acutely aging model rats were made by injecting D-gal into abdominal cavity continually. RT-PCR was used to detect expressions of p53 and Rb genes in the testes of rats and Western blotting to detect the expressions of phosphorylated Rb, p16, p19 and p21 proteins in the testes of rats. Results The expression of p53 and Rb in the testes of model rats were obviously higher than those in the normal control group (P0.01 ); The expressions of p16, p19 and p21 proteins increased obviously in the testes of model group rats compared with the normal group, while the phosphorylated Rb protein expression decreased significantly (P0.01 ). Conclusion The expressions of p53/Rb cellular transduction pathway related genes and proteins changed obviously in aging rat testes, which suggests that the blocking of p53/Rb cellular transduction pathway may play a certain role in the aging of testis.