Increase of nitric oxide production resulted in the development of oxidative-nitrosative stress that is considered to be an etiological cause of many diseases, including diabetes mellitus (DM). Recently, it was found that red blood cells (RBCs) are able to produce nitric oxide (NO), and due to the ability of hemoglobin to bind to nitric oxide, are the main depot of NO. Medicinal mushrooms are widely used in the correction and treatment of many diseases, including diabetes. Our previous results showed that Agaricus brasiliensis and Ganoderma lucidum have hypoglycemic effects and improve the functional state of RBCs. In this study, the influence of submerged cultivated mycelium powder (SCMP) of the abovementioned mushrooms on the red blood cells L-arginine/NO system of streptozotocin-induced DM rats was investigated. Wistar outbread white rats were used in the study. Streptozotocin was intraperitoneal injected one time at a dose of 50 mg/kg body weight. Mushroom preparations were orally administered at a dose of 1 g/kg/day for 14 days. We showed that administration of medicinal mushrooms SCMP to diabetic animals caused restoration of NO-synthase activity, normalized nitrite content (in case of A. brasiliensis), and led to nitrate growth. Therefore, treatment with mushroom mycelia normalizes the production of nitric oxide to physiological values.