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DOI: 10.18413/2313-8955-2018-4-4-0-7

Effects of ked peptide on connexin and sirutin expression in atherosclerosis and restinosis in elderly people

Background: Despite the wide range of application of the KED peptide, the molecular mechanisms of its action are still not fully understood. The aim of the study:To study the effect of the KED peptide on the expression of connexin and sirtuin in atherosclerosis and restenosis in the elderly. Materials and methods:For the study, the cell cultures obtained from the aorta without pathological changes, human atherosclerotic aorta, and human aorta during restenosis were used. Primary culture was isolated on Petri dishes (Sarstedt) treated with a solution of fibrinogen (Gibco), the subsequent cultivation was carried out in vials with a surface of 50 ml (Sarstedt 25 cm2). Cells were grown in 5 ml of culture medium per bottle and 3 ml of culture medium per Petri dish with a diameter of 3.5 cm, in 1 ml per well of a 24-well plate. In this work, the KED peptide was used in the form of a sterile solution, packaged in ampoules in a concentration of 100 μg in 1 ml. For immunocytochemical studies, the following primary monoclonal antibodies Connexin 37 (Dako, USA) and Sirtuin (Sirt)-1 (Abcam, UK) were used. To evaluate the results of immunocytochemical staining, a morphometric study was performed using a computer microscopic image analysis system including the Olympus BX40 microscope. Results:The expression of Connexin37 in the culture of the endothelium obtained from an elderly patient with atherosclerosis decreased by 3.6 times, and with stenosis – by 6.2 times compared with the norm. The KED peptide contributed to a 1.6-fold increase in Connexin 37 expression in cultures of endotheliocytes affected by atherosclerosis and 2.3 times in cultures affected by restenosis. The expression of Sirt1 in the endothelium in atherosclerosis and restenosis was 2 times lower compared with the norm. The KED peptide contributed to a 1.5-fold increase in the expression of Sirt1 in cultures of endotheliocytes affected by atherosclerosis and restenosis. Conclusion:High levels of Endothelin1 in the blood plasma are observed in atherosclerosis, ischemia and hypertension, therefore, the KED peptide, contributing to a decrease in the synthesis of this molecule, will help prevent the development of this cardiovascular pathology.

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