BACKGROUND: The three-year survival rate of patients with ischemic cardiomyopathy (ICMP) is 535 %; its pathogenesis is poorly understood. Violation of cytokine-dependent differentiation of monocytes/macrophages involved in atherogenesis may play an important role in the development of ICMP. AIMS: To characterize the disorders of monocytes subpopulation composition and mediators spectrum of blood in patients with coronary heart disease (CHD), associated with the development of ICMP. METHODS: A one-stage, clinical, controlled (case-control) study was conducted from February 2017 to December 2018. 45 patients with CHD (all men), who were in a cardiac surgery hospital, were examined before coronary bypass surgery: 19 people suffering from ICMP, and 26 people who do not suffer from ICMP, as well as 14 healthy men. In the blood of the examined individuals CD14++CD16-, CD14++CD16+, CD14+CD16++ and CD14+CD16- monocytes were determined with respect to all CD14-positive cells by flow cytometry, in blood plasma ― concentration of galectin 2 and 9, IL-4, IL-10, IFN-, M-CSF, HIF-1 by enzyme-linked immunosorbent assay (ELISA). RESULTS: The development of the ICMP accompanied by deficiency of HIF-1 and CD14+CD16++ monocytes (0.037 [0.020; 0.045] ng/ml, p = 0.019, and 5.05 [4.08; 6.58] %, p = 0.011) in combination with an excess of IL-10 (30.05 [24.75; 33.50] ng/ml, p = 0.042) in the blood. It is shown in blood of patients without ischemic cardiomyopathy the increase in the content of CD14++CD16+ cells and lack of CD14+CD16- monocytes (25.27 [15.78; 31.39] %, p = 0.038, and 2.68 [2.63; 4.09] %, p = 0.027) at normal concentration of IL-10 and HIF-1. In patients with CHD and ischemic cardiomyopathy and without ICMP in the blood the concentration of M-CSF and galectin-2 (2.00 [1.21; 3.24] pg/ml, p = 0.028, and 0.40 [0.12; 2.37] ng/ml, p = 0.004; 3.00 [1.90; 4.05] pg/ml, p = 0.003, and 3.20 [2.07; 4.00] ng/ml, p = 0.002, respectively) is reduced at normal content of galectin-9, and CD14++CD16- monocytes. IL-4 and IFN- in blood plasma are not determined (zero values). CONCLUSIONS: The development of ICMP is associated with excess of IL-10 and HIF-1 deficiency, which is accompanied by inhibition of CD14+CD16++ monocytes maturation.

Highlights. The features of subsets of monocytes in combination with the levels of desquamated endotheliocytes, endothelial damage and regeneration mediators and progenitor cell migration-enhancing factors in patients with coronary heart disease and with/without ischemic cardiomyopathy were analyzed. For the first time it was shown that in patients with ischemic cardiomyopathy, compared with CHD patients without cardiomyopathy, higher desquamation of the endothelium is associated with a deficiency of non-classical monocytes and reduced migration of progenitor endothelial cells (VEGFR2+-monocytes) with regenerative potential across the bone marrow due to a deficiency of the HIF-1α mediator in the blood.Background. The development of ischemic cardiomyopathy (ICM) is an understudied process, and one of its elements may be insufficient regeneration of blood vessels due to an imbalance of subsets of monocytes in the blood.Aim. To assess subsets of monocytes and desquamated endothelial cells in combination with endothelial damage and regeneration mediators in the blood of patients with coronary heart disease (CHD) and with/without ICM.Methods. The study included 30 patients with ICM, 22 patients with coronary heart disease without cardiomyopathy aged 55–69 years, and 18 healthy donors. In whole blood, the populations of CD45–CD146+ desquamated endothelial cells and progenitor endothelial cells related to CD14+VEGFR2+  monocytes, intermediate CD14++CD16+   and  non-classical  CD14+CD16++   monocytes  were  assessed  by flow cytometry  using  the  appropriate  monoclonal  antibodies  (BD  Biosciens, USA). In blood plasma, the levels of hypoxia-inducible factor HIF-1α, monocyte chemoattractant protein MCP-1 and matrix metalloproteinase MMP-9 were assessed by enzyme immunoassay. The results of the analysis were considered significant at p<0.05.Results. The number of progenitor and desquamated endothelial cells was increased in both groups of patients with coronary artery disease. At the same time, in patients with ICM, the number of progenitor endothelial cells did not reach the number noted in patients with CHD without cardiomyopathy, while the number of desquamated endothelial cells reached the number noted in CHD patients without cardiomyopathy. There was a deficiency of non-classical monocytes and HIF-1α in the blood of patients with ICM, and an excess of intermediate monocytes and MCP-1 was observed in CHD patients without cardiomyopathy. The concentration of MMP-9 in patients with CHD corresponded to the norm, regardless of the presence of ICM.Conclusion. In ICM, in contrast to CHD without cardiomyopathy, vascular damage is associated with a deficiency of nonclassical monocytes and reduced endothelial repair due to insufficient migration of progenitor endothelial cells across the bone marrow due to HIF-1α deficiency in the blood.

<p><strong>Background.</strong> Chronic heart failure frequently occurs against the background of ischemic heart disease (IHD), and has also been associated with the diagnosis of ischemic cardiomyopathy (ICMP). While there are significant commonalities with respect to underlying mechanisms, it would be most important to identify differential diagnostic markers to facilitate diagnosis and verification at the early stages of disease.</p><p><strong>Aim.</strong> The intent of this study was to perform a quantitative assessment of non-classical monocytes in patients diagnosed with IHD with or without ICMP and to identify any relationships between non-classical monocytes and plasma concentrations of proinflammatory (interleukin (IL)-1β, IL-6, tumour necrosis factor (TNF)-α) and anti-inflammatory (IL-4, IL-10, IL-13) cytokines.</p><p><strong>Methods.</strong> We examined 44 patients diagnosed with IHD, aged 49 - 63 years, with class II-III heart failure as per the New York Heart Association criteria. Of this group, 18 were also diagnosed with ICMP. The comparison group included 14 age-matched healthy controls. Percentages of non-classical (CD14+CD16+) monocytes in peripheral blood were determined by flow cytometry and the concentrations of IL-1β, IL-4, IL-6, IL-10, IL-13 and TNF-α were measured by enzyme-linked immunosorbent assay.</p><p><strong>Results.</strong> Amongst the patients diagnosed with ICMP, the concentration of IL-10 in blood plasma was higher than in those without this diagnosis, detected at 30.05 pg/ml (range 24.75 - 33.50 pg/ml, р=0.0412); the percentage of non-classical monocytes was lower, at 5.05% (range 4.08 - 6.58%, р=0.0094). Amongst patients diagnosed with IHD without ICMP, these parameters were within normal limits. The plasma concentrations of IL-1β, IL-6 and IL-13 in both groups were comparable to values obtained from healthy donors and IL-4 was undetectable throughout. Amongst patients diagnosed with ICMP, plasma TNF-α was detected at elevated levels that were comparable to those determined for patients diagnosed with IHD without ICMP.</p><p><strong>Conclusion.</strong> Imbalance of cytokines in blood in patients with coronary artery disease is characterized by excess TNF-α, absence of IL-4, and normal levels of IL-1β, IL-6 and IL-13, regardless of the type of ischemic myocardial injury. In patients with ICMP, we detected an overall decrease in the fraction of non-classical monocytes in association with an increase in plasma IL-10; these features were not detected in patients diagnosed with IHD without ICMP. As such, these biological responses may be used for timely diagnosis of ICMP.</p><p>Received 27 May 2019. Revised 7 October 2019. Accepted 30 October 2019.</p><p><strong>Funding:</strong> The work is supported by grants of the Russian Foundation for Basic Research No. 18-015-00160\19 and the President of the Russian Federation No. НШ-2690.2018.7 and No. МД-2788.2019.7.</p><p><strong>Conflict of interest:</strong> Authors declare no conflict of interest.</p><p><strong>Author contributions</strong></p><p><strong></strong>Conception and study design: V.M. Shipulin, S.P. Chumakova</p><p>Data collection and analysis: V.M. Shipulin, S.P. Chumakova, O.I. Urazova, M.V. Vins, V.V. Novickiy</p><p>Literature review: S.P. Chumakova, D.A. Pogonchenkova, A.S. Pryakhin</p><p>Drafting the article: S.P. Chumakova, D.A. Pogonchenkova, O.I. Urazova</p><p>Critical revision of the article: V.M. Shipulin, V.V. Novickiy</p><p>Statistical analysis: S.P. Chumakova</p><p>Final approval of the version to be published: V.M. Shipulin, S.P. Chumakova, D.A. Pogonchenkova, O.I. Urazova, M.V. Vins, A.S. Pryakhin, V.V. Novickiy</p>

Цель исследования оценить соотношение фракций классических, промежуточных, неклассичес- ких и переходных моноцитов во взаимосвязи с концентрацией интерлейкинов (IL) 4 и 10 в крови у больных ишемической кардиомиопатией. Материал и методы. Обследованы 18 больных ишемической кардиомиопатией (17 мужчин и 1 женщина) в возрасте 4766 лет с недостаточностью кровообращения IIIII функционального класса по классификации сердечной недостаточности Нью-Йоркской ассоциации кардиологов. Группу контроля составили 14 практически здоровых доноров, сопоставимых по полу и возрасту с больными ишемической кардиомиопатией, не имеющих каких-либо заболеваний сердечно-сосудистой системы, а также других систем органов в стадии обострения. У больных ишемической кардиомиопатией в крови оценивали относительное содержание классических (CD14CD16), промежуточных (CD14CD16), неклассических (CD14CD16) и переходных (CD14CD16) моноцитов методом проточной цитометрии и концентрацию IL-4 и IL-10 методом иммуноферментного анализа. Результаты. Уровень неклассических моноцитов в крови у больных ишемической кардиомиопатией оказался существенно ниже нормы (5,05 4,08 6,58 и 10,07 9,34 13,84 соответственно, р0,008), как и концентрация IL-4 (0,02 0 0,04 пг/мл и 0,15 0,05 0,63 пг/мл соответственно, р0,039). Содержание IL-10 в крови было повышенным (30,05 24,75 33,50 пг/мл, р0,042) и негативно коррелировало с числом неклассических моноцитов (r0,65 p0,02). Количество классических моноцитов в крови у пациентов имело тенденцию к снижению, а доля промежуточных моноцитов, напротив, была несколько выше, чем у здоровых лиц. Относительное содержание переходных моноцитов в крови оказалось сопоставимым с показателями у здоровых доноров. Заключение. Субпопуляционный состав моноцитов крови у больных ишемической кардио- миопатией характеризуется дефицитом фракции неклассических моноцитов, обладающих протек- тивными свойствами в отношении эндотелия, и IL-4, при избытке IL-10 и некотором увеличении количества промежуточных клеток, обладающих способностью к кооперации с Т-лимфоцитами, что предрасполагает к распространенному атероматозу мелких коронарных артерий и диффузному гипоксическому поражению миокарда при ишемической кардиомиопатии.