A streptozotocin‐nicotinamide‐induced T2DM model was generated in 50 random rats using the method reported by Bayrasheva et al. 9 The remaining 10 animals were in the intact control group. We used a high‐fat diet for this model to create the metabolic changes associated with T2DM. 9 These 50 animals were transferred for 5 weeks to a high‐fat diet with the addition of beef fat to the standard feed. The total caloric content of the food was 450 kcal/100 g (20%–fat, 48%–carbohydrates and 20%–protein). 9 These rats were without food for 12 hours after a period of high‐fat diet. We injected a nicotinamide solution (230 mg/kg) intraperitoneal, then 15 minutes later we injected a streptozotocin solution (65 mg/kg) intraperitoneal. 10 Nicotinamide (Herba Hemosan, Austria) was previously dissolved in 0.9% sodium chloride solution. Streptozotocin (Sigma Aldrich) was dissolved in a freshly prepared sodium citrate buffer. 11 We injected 0.9% sodium chloride solution to 10 intact animals during the simulation of T2DM.

The development of experimental type 2 diabetes in rats is confirmed by monitoring the level of fasting glycemia (on an empty stomach at 9:00 am, after 8–12 h of starvation), followed by the oral glucose tolerance test (OGTT) with oral or parenteral administration of glucose solution. 12

Diagnostic criteria for T2DM included fasting blood glucose level 9–14 mmol/L (OneTouch Select glucometer, LifeScan Inc.) and more than 2‐fold increase in the calculated area under the glucose curve based on OGTT results in comparison with intact animals. 12

Experimental studies were conducted at the Federal State Budget Scientific Institution ‘Institute of Experimental Medicine’ in cooperation with the staff of the Laboratory of Chemistry and Pharmacology of Medicine, following the ‘Guidelines for the Care and Use of Laboratory Animals’ and ‘A guide to the experimental (preclinical) study of new pharmacological substances’, in accordance with the principles of humanity, European Directives (86/609/EEC) and Helsinki Declaration.

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