Author contributions: Zykov VA, Tuchina TP and Lebedev DA performed the experiments and wrote the manuscript; Babenko AY, Kuleshova EV and Grineva EN performed the literature review and suggested the study concept; Krylova IB and Bayramov AA developed the experimental design and performed data analysis; Galagudza MM provided scientific consulting, coordinated experimental parts, and edited the manuscript. Supported by Russian Science Foundation, No. Correspondence to: Alina Y Babenko, DSc, MD, PhD, Doctor, Research Scientist, Institute of Endocrinology, Almazov National Medical Research Centre, Akkuratova St., 2, St-Petersburg 197341, Russia. Telephone: +7-9 Fax: +7-8. METHODS Type 2 diabetes mellitus (T2DM) was induced in male Wistar rats with streptozotocin (65 mg/kg) and verified using an oral glucose tolerance test.

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After anesthesia, the left coronary artery was occluded for 40 min followed by 80 min reperfusion. Blood glucose level was measured during surgery. Rats were randomized into six groups as follows: (1) control rats; (2) insulin (0.1 U/kg) treated rats prior to ischemia; (3) insulin (0.1 U/kg) treated rats at reperfusion; (4) GLP-1a (140 mg/kg) treated rats prior to ischemia; (5) GLP-1a (140 mg/kg) treated rats at reperfusion; and (6) rats treated with GLP-1a (140 mg/kg) prior to ischemia plus insulin (0.1 U/kg) at reperfusion. Myocardial area at risk and infarct size was measured planimetrically using Evans blue and triphenyltetrazolium chloride staining, respectively. RESULTS There was no significant difference in the myocardial area at risk among groups. Insulin treatment before ischemia resulted in a significant increase in infarct size (34.7% ± 3.4% vs 18.6% ± 3.1% in the control rats, P. Core tip: In addition to their glucose-lowering effects, glucagon-like peptide-1 analogs (GLP-1a) were shown to exhibit cardioprotective effects.

However, the optimal protocol of GLP-1a administration for infarct size reduction has not been determined yet. Additionally, it is important to investigate the effects of GLP-1a combined with other antidiabetic drugs on myocardial infarct size. Thus, we evaluated the effects of GLP-1a with and without insulin on infarct size in rats with type 2 diabetes mellitus. We found that GLP-1a administration prior to ischemia resulted in significant infarct size reduction. Infarct size reduction was maximal in rats treated with GLP-1a before ischemia plus insulin at reperfusion.

INTRODUCTION Type 2 diabetes mellitus (T2DM) is considered a risk factor for cardiovascular diseases with an approximately three-fold increased risk of myocardial infarction (MI). Normalizing glucose variability can prevent future cardiovascular complications. Safe blood glucose levels during MI (10 mmol/L, ideally. Animals Seventy male Wistar rats were used in this study.

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Both neonatal STZ-induced T2DM and MI were induced in these rats. 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, in accordance with the “Guidelines for the Care and Use of Laboratory Animals” and “A guide to experimental (preclinical) study of new pharmacological substances,” with observance of the principles of humanity, European Directives (86/609/EEC), and Helsinki Declaration.

T2DM model We used the streptozotocin(STZ)-nicotinamide model of diabetes. Induction of diabetes in rats was carried out by a single intraperitoneal injection of STZ at 65 mg/kg, dissolved in citrate buffer (pH 5.5). For selection of rats to be used in the study, blood glucose level was measured at the age of 3 mo, followed by an oral glucose tolerance test after administration of 40% w/v glucose solution at a dose of 3 g/kg. Diagnostic criteria for T2DM included fasting blood glucose levels from 7 to 14 mmol/L (OneTouch Select glucometer, LifeScan Inc., Milpitas, CA, United States) and two-fold increase in the area under the glucose curve of the oral glucose tolerance test, compared with that in the control group[,].

Myocardial ischemia-reperfusion model Rats were anesthetized using chloral hydrate solution (400 mg/kg), tracheotomized, and ventilated (SAR-830P, Stoelting, United States) using room air, with a tidal volume of 2 mL/100 g and a rate of approximately 60 breaths/min. The core body temperature was maintained at 37 ± 0.5 °C using a feedback-controlled heating pad (TCAT-2LV controller, Physiotemp Instruments Inc., Clifton, NJ, United States). The left carotid artery and right femoral vein were cannulated for measurement of the mean arterial pressure (MAP) and maintenance of anesthesia, respectively. Lead II of the electrocardiograph was monitored for determination of the heart rate (HR) and arrhythmias. After a 10 min stabilization, a left thoracotomy was performed. A 6-0 polypropylene thread was placed around a prominent branch of the left coronary artery, and the ends were passed through a polyethylene tube as an occluder. Exclusion criteria were MAP.