Chronic heart failure (HF) remains a leading cause of cardiovascular (CV) mortality and morbidity worldwide. The aim of the study was to investigate whether the pattern of angiogenic endothelial progenitor cells (EPCs) and apoptotic endothelial cell-derived microparticles (EMPs) would be able to differentiate HF with reduced (HFrEF) and preserved (HFpEF) ejection fraction. One hundred sixty four chronic HF subjects met inclusion criteria. Patients with global left ventricular ejection fraction ≥ 50% were categorized as the HFpEF group (n = 79) and those with ≤ 45% as the HFrEF group (n = 85). Therefore, to compare the circulating levels of biological markers 35 control subjects without HF were included in the study. All control individuals were age- and sex-matched chronic HF patients. The serum level of biomarkers was measured at baseline. The flow cytometric technique was used for predictably distinguishing circulating cell subsets depending on expression of CD45, CD34, CD14, Tie-2, and CD309 antigens and determining endothelial cell-derived microparticles. CD31(+)/annexin V(+) was defined as apoptotic endothelial cell-derived MPs, MPs labeled for CD105(+) or CD62E(+) were determined as MPs produced due to activation of endothelial cells. In multivariate logistic regression model T2DM (R(2) = 0.26; P = 0.001), obesity (R(2) = 0.22; P = 0.001), previous MI (R(2) = 0.17; P = 0.012), galectin-3 (R(2) = 0.67; P = 0.012), CD31(+)/annexin V(+) EMPs (R(2) = 0.11; P = 0.001), NT-proBNP (R(2) = 0.11; P = 0.046), CD14(+) CD309(+) cells (R(2) = 0.058; P = 0.001), and CD14(+) СD309(+) Tie-2(+) cells (R(2) = 0.044; P = 0.028) were found as independent predictors of HFpEF. Using multivariate Cox-regression analysis adjusted etiology (previous myocardial infarction), cardiovascular risk factors (obesity, type 2 diabetes mellitus) we found that NT-proBNP (OR 1.08; 95% CI = 1.03-1.12; P = 0.001) and CD31(+)/annexin V(+) EMPs to CD14(+) CD309(+) cell ratio (OR 1.06; 95% CI = 1.02-1.11; P = 0.02) were independent predictors for HFpEF. We found that CD31(+)/annexin V(+) EMPs to CD14(+) CD309(+) cell ratio added to NT-proBNP, clinical data, and cardiovascular risk factors has exhibited the best discriminate value and higher reliability to predict HFpEF compared with NT-proBNP and clinical data/cardiovascular risk factors alone.

The study aim was to evaluate whether circulating microparticles with apoptotic or non-apoptotic phenotypes are useful for risk assessment of 3-year cumulative fatal and non-fatal cardiovascular events in CHF patients. The incidence of fatal and non-fatal cardiovascular events, as well as the frequency of occurrence of death from any cause in a cohort of 388 patients with CHF during 3 years of observation was studied prospectively. Circulating levels of NT-pro brain natriuretic peptide (NT-pro-BNP), high-sensitivity C-reactive protein (hs-CRP), and endothelial apoptotic microparticles (EMPs) were measured at baseline. Median follow-up was 2.32 years (IQR = 1.8-3.1). During follow-up, 110 cardiovascular events (including 43 fatal cases) were determined. Additionally, 74 subjects were hospitalized repetitively due to worsening CHF and also 16 subjects were readmitted in the hospital due to other cardiovascular reasons. In the univariate logistic regression analysis, the main factors independently related with cumulative endpoints were creatinine, fasting glucose, HbA1c, total cholesterol, uric acid, various types of EPMs, NT-pro-BNP, hs-CRP, NYHA class, decreased left ventricular ejection fraction (LVEF) less 45%, and type 2 diabetes mellitus. In multivariate model NYHA class, decreased LVEF (less 45%), NT-pro-BNP, hs-CRP, CD144 +/CD31 +/annexin V + EMPs, and CD31 +/annexin V + EMPs remained statistically significant for cumulative endpoint. Adding of CD144 +/CD31 +/annexin V + EMCs and CD31 +/annexin V + EMCs to the standard ABC model may improve the relative IDI for cumulative endpoint by 11.4% and 10.5% respectively. Apoptotic phenotype of circulating microparticles may relate 3-year combined clinical outcomes in CHF patients.

Background: Accelerating atherosclerosis in type 2 diabetes mellitus (T2DM) patients may relate to imbalance between pattern of microparticles (MPs), which are frequently involved in repair of vasculature, tissue injury, inflammation and thrombosis. The aim of the study was to investigate the pattern of circulating MPs in T2DM patients with asymptomatic atherosclerosis. Methods: A total of 103 patients with T2DM (54 subjects without documented coronary atherosclerosis and 49 patients with angiographic evidence of asymptomatic coronary atherosclerosis) who were underwent a contrast-enhanced multispiral computed tomography angiography and 35 healthy volunteers were enrolled in the study. To determine circulating biomarkers, blood samples were collected at baseline. MPs were labelled and characterized by flow cytometry. Results: There were no significant differences between healthy volunteers and T2DM patients in circulating numbers of MPs labelled as CD41a+, CD64+, CD144+, CD144+/CD31+, Annexin V+, CD144+/annexin V+ and CD144+/CD31+/annexin V+. However, lower number of MPs with immune phenotypes CD62E+, CD105E+ and higher numbers of CD31+/annexin V+ MPs were reported in T2DM patients when compared with healthy volunteers. Therefore, we found an increased level of circulating CD41a+ MPs, CD144+/CD31+ MPs, CD31+/annexin V+ MPs, and decreased level of CD62E+ MPs in T2DM patients with asymptomatic coronary atherosclerosis in comparison with those who had no asymptomatic atherosclerosis. Using multivariate log regression analysis, BMI (odds ratio [OR] = 1.04, p = 0.001), LDL-C (OR = 1.05, p = 0.046), hs-CRP (OR = 1.07, p = 0.044), osteoprotegerin (OR = 1.07, p = 0.026), CD62E+ MPs (OR = 1.07, p = 0.001) and CD31+/annexin V+ MPs (OR = 1.12, p = 0.003) were determined independent predictive factors of asymptomatic atherosclerosis in T2DM patients. Conclusion: Circulating levels of MP originated from apoptotic endothelial cell-derived were significantly increased in diabetic patients as compared with normal subjects, but level of activated endothelial cell-derived MPs was lower than in healthy volunteers. Among T2DM patients, an increased level of CD31+/annexin V+ MPs and decreased CD62E+ MPs were significantly associated with asymptomatic atherosclerosis.