Study of the immobilized subtilisins influence on coronary blood fl ow of an isolated rat heart

Introduction. In clinical medicine, some phenomenon has been noted: in many patients with anginal attacks during coronary angiography, significant obstructive processes in the coronary arteries are not detected. There is impaired coronary microcirculation due to dysfunction of endothelial and smooth muscle cells. This circumstance initiates the metasearch for new drugs for pharmacological correction of the vasomotor status of coronary vessels in order to eliminate myocardial ischemia.

Aim. To study the parameters of the coronary blood flow during the perfusion of an isolated heart with immobilized subtilisins (IS) in different dosages.

Materials and methods. The effect of IS on coronary blood fl ow was studied using the Langendorff-perfused rat heart. The experiments were carried out on 50 male Wistar rats. Animals were divided into 5 groups: control (hearts perfused only with Krebs-Henseleit solution) and 4 experimental (1st — perfusion with IS at a concentration of 170 U/l; 2nd — perfusion with IS at a concentration of 340 U/l; 3rd — perfusion with IS at a concentration of 500 U/l; 4th — perfusion with IS at a concentration of 1000 U/l).

Results. An increase in coronary blood flow was observed during perfusion with IS at concentrations of 170 and 340 U/l. With perfusion of IS at a concentration of 340 U/l, the vasodilating effect was significantly expressed. With perfusion at a concentration of 500 and 1000 U/l, a decrease in coronary flow was noted, which is most likely caused by vasoconstriction.

Conclusion. When the heart is perfused with IS solution, the phenomenon of vasomotor activity of the coronary vessels is observed. The vasomotor effect caused by IS has a dose-dependent and multidirectional effect and is realized, most likely, through an allosteric effect on the natural mechanism of regulation of the coronary vessels tone.

1. Vancheri F., Longo G., Vancheri S., Henein M. (2020). Coronary microvascular dysfunction. J. Clin. Med., 9 (9): 2880. doi: 10.3390/jcm9092880.

2. Cziráki A., Lenkey Z., Sulyok E., Szokodi I., Koller A. (2020). L-arginine-nitric oxide-asymmetric dimethylarginine pathway and the coronary circulation: translation of basic science results to clinical practice. Front. Pharmacol., 11: 569914. doi: 10.3389/fphar.2020.569914.

3. Goodwill A.G., Dick G.M., Kiel A.M., Tune J.D. (2017). Regulation of coronary blood flow. Compr. Physiol., 7 (2), 321–382. doi: 10.1002/cphy.c160016.

4. Tune J.D. (2014). Coronary circulation. Colloquium Series on Integrated Systems Physiology: from Molecule to Function to Disease, 6 (3), 1–189. doi: 10.4199/C00111ED1V01Y201406ISP054.

5. Duncker D.J., Bache R.J. (2008). Regulation of coronary blood fl ow during exercise. Physiol. Rev., 88 (3), 1009–1086. doi: 10.1152/physrev.00045.2006.

6. Konst R.E., Guzik T.J., Kaski J.C., Maas A.H.E.M., Elias-Smale S.E. (2020). The pathogenic role of coronary microvascular dysfunction in the setting of other cardiac or systemic conditions. Cardiovasc. Res., 116 (4), 817–828. doi: 10.1093/cvr/cvaa009.

7. Deussen A., Ohanyan V., Jannasch A., Yin L., Chilian W. (2012). Mechanisms of metabolic coronary flow regulation. J. Mol. Cell. Cardiol., 52 (4), 794–801. doi: 10.1016/j.yjmcc.2011.10.001.

8. Madonov P.G., Kinsht D.N., Ershov K.I., Shilova M.A., Solov’ev O.N. (2015). Clinical experience of the new drug, Trombovazim® in vascular surgery. Angiology and Vascular Surgery, 21 (1), 99–104. In Russ.

9. Madonov P.G., Mishenina S.V., Kinsht D.N., Kikhtenko N.V. (2016). Targeted pharmacodynamics of subtilisins. The Siberian Scientifi c Medical Journal, 36 (4), 15–23.

10. Pharmaceutical composition with thrombolytic, antiinflammatory and cytoprotective properties: RF Patent No. 2213557 / А.V. Artamonov, E.I. Vereshchagin, O. V. Grishin, A.V. Troitsky. Published: 10.10.2003.

11. Minasyan S.M., Galagudza M.M., Sonin D.L. et al. (2009). Isolated rat heart perfusion technique. Region al Blood Circulation and Microcirculation, 8, 4 (32), 54–59. In Russ.

12. Bell R.M., Mocanu M.M., Yellon D.M. (2011). Retrograde heart perfusion: the Langendorff technique of isolated heart perfusion. J. Mol. Cell. Cardiol., 50 (6), 940–950.

13. Watanabe M., Okada T. (2018). Langendorff perfusion method as an ex vivo model to evaluate heart function in rats. Methods Mol. Biol., 1816, 107–116. doi: 10.1007/978-1-4939-8597-5_8.

14. On the approval of the Rules of Good Laboratory Practice: Order of the Ministry of Health of the Russian Federation dated 01.04.2016 No. 199н. Retrieved on June 8, 2021 from http://publication.pravo.gov.ru/Document/View/0001201608160001?index=0&rangeSize=1.