Combined Correction of Experimental Critical Ischemia of the Lower Extremities in a Rat Model

Elvin E. Feyziev, Boris S. Sukovatykh, Alexander S. Belous, Maria A. Zatolokina, Elena V. Trubnikova, Ekaterina S. Mishina

International Journal of Biomedicine. 2021;11(3):372-375.
DOI: 10.21103/Article11(3)_OA16
Originally published September 9, 2021


The purpose of our research was to study the effectiveness of correcting experimental critical ischemia (CI) of the lower extremities with a combination of udenafil, simvastatin, and autologous bone marrow mononuclear cells (ABMMC).
Methods and Results: The experiments were carried out on 24-month-old Wistar rats, weighing 220–250g. The animals were randomized by sex and weight. Groups were formed according to the manipulations carried out during the operations. The animals were divided into 7 groups, each with 20 animals: Group 1 included intact animals; Group 2 - falsely operated animals; Group 3 (control group) - animals with simulated CI without treatment; Group 4- animals with CI and monotherapy with udenafil (daily oral administration of 8.6mg/kg for 28 days); Group 5 - animals with CI and simvastatin monotherapy  (daily oral administration of 1.71mg/kg for 28 days); Group 6 - animals with CI and monotherapy with ABMMC (parenterally, once on Day 7 after modeling CI, 50μl at 4 points and, paravasally, above the inguinal ligament in the area where the lateral artery leaves the artery enveloping the femur from the internal iliac artery; in the area of the superficial artery that bends around the iliac bone under the inguinal ligament; into the area of origin of the muscular branch of the femoral artery r. muscularis, the place of attachment of the comb and long adductor muscles of the thigh; in the upper third of the gastrocnemius muscle]); Group 7 - animals with CI and combination therapy (udenafil and simvastatin drugs were administered intragastrically 0.86 mg/kg, once a day, for 7 days) and one-time parenteral administration of ABMMC, according to the same scheme as in Group 6. On Days 21 and 28 of the experiment, the level of blood microcirculation was determined in the muscles of the rat leg; for this, laser Doppler flowmetry was used. For further morphometric assessment of the leg muscles, they were removed. Preparations for morphometric analysis were prepared according to the standard technique with Van Gieson staining, as well as H&E. Our study demonstrated the effectiveness of combination therapy with udenafil, simvastatin, and ABMMC to correct critical lower limb ischemia in rats. The severity of morphological changes on the background of this combination was minimal, compared to the findings of other study groups, and the level of blood microcirculation in the ischemic zone on Day 28 was, significantly, 1.9 times higher than in animals of the control group. The results obtained allow us to recommend the use of the investigated combination (udenafil+simvastatin+ABMMC) for the treatment of patients with critical limb ischemia, both in outpatient and inpatient practice.

critical ischemia • microcirculation • autologous bone marrow mononuclear cells

1. Sumin AN, Medvedeva JuD, Shheglova AV, Barbarash LS. [Predictors of adverse outcomes in patients with atherosclerosis obliterans of the lower limb arteries]. Cardiology and Cardiovascular Surgery. 2020;13(1):41-7. [Article in Russian].
2. Emeljanov SG, Korenevskij NA, Bykov AV. [Predicting the severity of the ischemic process in the heart, brain and lower extremities based on fuzzy models]. Biomedical Radio Eeectronics. 2016;9:4–9. [Article in Russian].
3. Genkel VV, Salashenko AO, Shamaeva TN, Sumerkina VA, Nikushkina KV, Shaposhnik II. [Atherosclerosis of peripheral arteries in patients with coronary artery disease and type 2 diabetes mellitus]. Ter Arkh. 2019 Oct 15;91(10):54-62. doi: 10.26442/00403660.2019.10.000106.  [Article in Russian].
4. Bogachev VYu, Boldin BV, Rodionov SV, Turkin PYu. Conservative treatment of chronic obliterating diseases of lower limb arteries. Ambulatornaya Khirurgiya = Ambulatory Surgery (Russia). 2018;(1-2):14-19. [Article in Russian].
5. Kondratyeva LV. [Barriers to statin therapy in patients with type 2 diabetes mellitus and ways to overcome them]. Russian Medical Journal. 2017;22:1621-1628. [Article in Russian].
6. Arutyunov GP, Boytsov SA, Voevoda MI, Gurevich VS, Drapkina OM, Kukharchuk VV, et al. [Correction of hypertriglyceridemia in order to reduce the residual risk in diseases caused by atherosclerosis]. Conclusion of the Council of Experts of the Russian Society of Cardiology, the Russian Scientific Medical Society of Physicians, the Eurasian Association of Physicians, the National Society for the Study of Atherosclerosis, the Russian Association of Endocrinologists and the National Research League of Cardiological Genetics. Rational Pharmacotherapy in Cardiology. 2019;15(2):282-8. [Article in Russian].
7. Osipova OS, Saaia SB, Karpenko AA, Zakiian SM. Problemy i perspektivy kletochnoĭ terapii kriticheskoĭ ishemii nizhnikh konechnosteĭ [Problems and prospects of cell therapy for critical ischaemia of lower limbs]. Angiol Sosud Khir. 2020;26(2):23-33. doi: 10.33529/ANGIO2020220. [Article in Russian].
8. Feyziev EE, Belous AS, Sukovatykh BS, Trubnikova EV. Kursk State University, patent holder. A method for modeling critical ischemia of the lower extremities in experimental rat animals. RF patent, RU 2734158 dated 03/18/2020.
9. Böyum A. Separation of leukocytes from blood and bone marrow. Introduction. Scand J Clin Lab Invest Suppl. 1968;97:7. 
10. Kolesnik IM, Lazarenko VA, Pokrovsky MV. [Effect of pharmacological preconditioning with sildenafil and vardenafil on the state of the microvasculature in ischemic skeletal muscle]. Kursk Scientific and Practical Bulletin "Man and His Health." 2015;(1):83-6. [Article in Russian].

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Received July 30, 2021.
Accepted August 30, 2021.
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