Background: In both human and rat tissues, the peptide hormone spexin is expressed. Some studies examined its relation to obesity and diabetes. Aim: To investigate the impact of spexin on testicular alterations in type 2 diabetic rats, both functionally and histologically, and potential underlying processes. Material and Methods: Four equal groups of twenty-four adult rats were used: control, type 2 diabetes group, type 2 diabetic group treated with metformin, and type 2 diabetic group treated with spexin. Rats were given a high-fat diet for eight weeks and injected single dose of streptozotocin (40 mg/kg) in the 4th week. These rats had developed type 2 diabetes. Normal diet and vehicle were provided to control rats. Treatment groups (III & IV) received either oral metformin (300 mg/kg/day) or intraperitoneal spexin (35 μg/kg/day) for four weeks, starting from the fifth week and ending at the end of the eighth week. Metabolic profile, some inflammatory, oxidative stress markers and serum spexin levels were assessed at the end of experiment. Histological and immunohistochemical assessment of the testis were investigated. Results: Diabetic groups showed a significant deterioration in metabolic profile, histological structure of the testis, epidydimal sperm count and motility, some inflammatory oxidative stress markers while testicular superoxide dismutase and serum spexin were significantly reduced. These alterations were significantly reversed in groups treated with spexin and metformin. Conclusion: Both Spexin and metformin have almost the same protective effect by attenuating the testicular histological and metabolic alterations caused by type 2 diabetes.