In the 21st century, the use of natural resources in the food and medicine industries has emerged as a significant global focus, primarily due to their low toxicity and high specificity in modulating the immune system. Among these resources, mushrooms have been extensively studied for their diverse bioactive properties, including antibacterial, antifungal, antioxidant, antiviral, antitumor, cytostatic, immunosuppressive, antiallergic, antiatherogenic, hypoglycemic, anti-inflammatory, and hepatoprotective activities. This study aimed to investigate the effects of a diet supplemented with Lentinus edodes on the serum enzyme profiles of male albino Wistar rats. The results indicated that the group of rats receiving a diet with 10% L. edodes (Group D) exhibited significant alterations in their blood serum profiles compared to the control groups. These findings suggest that dietary supplementation with L. edodes may influence metabolic and immune responses in male albino Wistar rats, warranting further investigation into its therapeutic potential.