Objective To investigated the effects of obesity on the structure of seminiferous tubules, autophagy and apoptosis-related proteins in testes tissue, and the effect of exercise on autophagy and apoptosis in testis and regulatory mechanism.Methods Fifty 6-week-old SD male rats were randomly divided into the standard diet group (SD, n=20) and the high-fat diet group (HFD, n=30). HFD group was fed 8 weeks to establish an obesity model of rats, and 20 rats were randomly selected to exercise intervention. Rats in SD and HFD groups were randomly divided into normal diet control group (CC), normal diet+exercise group (CE), high-fat diet control group (OC), and high-fat diet+exercise group (OE), 10 rats in each group. The rats in the CE and OE groups underwent moderate intensity treadmill training for 60 min/d, 5 d/week. CC and OC groups maintained the same feeding conditions. After 48 h of the last exercise, the rats were anesthetized intraperitoneally and weighed, the testes were taken from both sides of the rats, weighed and the testicular index was calculated. Paraffin sections of the testes were prepared to discover the testicular tissue structure by HE staining. Western blot was used to detect the protein expression of p62, LC3II, LC3I, BCL-2, Bax, and AMPK in testicular tissue, and the ratio of LC3II/LC3I was calculated. Immunofluorescence was used to detect LC3 and BCL-2 protein expression positions in the testis.Results The testicular index was reduced; HE staining revealed a significant decrease in the diameter of the seminiferous tubules (P<0.01), spermatocytes, and lipid droplet deposition in the testicular tissue; the protein expression of p62 and Bax were significantly increased (P<0.01), AMPK, LC3II/LC3I ratio, and BCL-2 were significantly decreased (P<0.01), and the number of LC3 proteins on sperm cells were significantly reduced in the testis tissue of rats in the OC group compared with the CC group. After exercise intervention, the testicular index was increased in the OE group; HE staining showed that the diameter of seminiferous tubules (P<0.01) and the number of spermatogonia (P<0.05) were significantly increased, and the structure of seminiferous tubules was improved; the protein expression of p62 and Bax were significantly decreased (P<0.01), AMPK, LC3II/LC3I ratio, and BCL-2 were significantly increased (P<0.01); the level of BCL-2 proteins on spermatocytes and sperm cells increased significantly in the OE group compared with OC group. AMPK was strongly correlated with p62, LC3II/LC3I, and Bax in all rat testicular tissue.Conclusion High-fat diet-induced obesity inhibits autophagy and promotes apoptosis in rat testes; 8 weeks of moderate-intensity exercise may activate autophagy and inhibit apoptosis in testis tissue via AMPK, correcting the adverse changes in spermatogenesis caused by obesity.