Angiogenesis, Metabolism, Endothelial And Platelet Markers In Diabetes And Cardiovascular Disease

Introduction: Diabetes is a leading risk factor for cardiovascular disease (CVD), the pathophysiology of both being linked to metabolic, endothelial, renal, angiogenic and platelet abnormalities. We hypothesised that abnormalities in these systems are more adverse in those whose CVD is compounded by diabetes, compared to those with diabetes or CVD alone.Materials and methods: Serum or plasma from 66 patients with diabetes alone, 76 with CVD alone, and 70 with both diabetes and CVD i.e. diabetic cardiovascular disease, was probed for markers of angiogenesis [angiopoietin 1 and 2, vascular endothelial growth factor (VEGF) and endoglin], metabolic [sRAGE, leptin, lipocalin-2, interleukin-8, and cystatin-C], the endothelium (von Willebrand factor, endothelial microparticles and soluble E selectin]), and the platelet (platelet microparticles and soluble P selectin) by ELISA, Luminex or flow cytometry.Results: VEGF (p=0.04), von Willebrand factor (p=0.001) and endothelial microparticles (p=0.042) were all higher in diabetic cardiovascular disease than in diabetes alone and cardiovascular disease alone. Soluble E selectin was higher in diabetic cardiovascular disease than in diabetes alone (p=0.045), whilst cystatin-C (p=0.004) and soluble P selectin (p<0.001) were higher in diabetes and diabetic cardiovascular disease than in cardiovascular disease alone. There were no differences in angiopoietin 1 or 2, endoglin, sRAGE, leptin, lipocalin-2, or interleukin-8.Conclusion: Angiopoietin 1 or 2, endoglin, sRAGE, leptin, lipocalin-2, interleukin-8, and cystatin-c cannot differentiate diabetes from cardiovascular disease, or both conditions combined. Our data point to a more adverse endothelial (von Willebrand factor, endothelial microparticles), and angiogenic profile (VEGF) in those with diabetic cardiovascular disease, supporting the view that this group should be targeted more aggressively.

Effects of modified-Paleo and moderate-carbohydrate diets on body composition, serum levels of hepatokines and adipocytokines, and flow cytometric analysis of endothelial microparticles in adults with metabolic syndrome: a study protocol for a randomized clinical trial

Background Metabolic syndrome is a combination of metabolic risk factors causing a pathological condition that increases the risk of non-communicable diseases, such as diabetes and cardiovascular diseases. A variety of dietary approaches have been examined to halt this rapid trend; however, the effects of modified-Paleo diet and medium-carbohydrate diet on inflammation, adipokines, hepatokines, and the profile of endothelial microparticles in individuals with metabolic syndrome have not been investigated in detail. The present study is designed to examine the effect of modified-Paleo and moderate-carbohydrate diet with two delivery modes: “fixed diet plan” vs “calorie counting” on weight, body composition, serum levels of some hepatokines and adipocytokines, and flow cytometric analysis of endothelial microparticles in adults with metabolic syndrome. Methods Eighty metabolic syndrome patients will be recruited in this study. They will be randomly allocated to one of the following 4 groups: (1) receiving a modified-Paleo diet with calorie counting, (2) receiving a modified-Paleo diet with a fixed diet plan, (3) receiving a medium-carbohydrate diet with calorie counting, and (4) receiving a medium-carbohydrate diet with a fixed diet plan for 10 weeks. Weight, height, waist circumference, and body composition will be assessed at the study baseline and at the end of the trial. Serum insulin, asprosin, chemerin, FGF-21, CTRP-1, PYY, ghrelin, plasma EMPs (CD31+/CD42b− and CD144+/CD42b−), lipid profile, glycemic indices, hs-CRP, leptin, vitamin C, creatinine and satiety, hunger, fullness, and desire to eat (via visual analog scales) will be measured at the study baseline and at the end of the trial. Insulin resistance and insulin sensitivity will be determined using the HOMA-IR and QUICKI equations. Discussion To the best of our knowledge, this is the first randomized controlled trial that will determine the effect of modified-Paleo and moderate-carbohydrate diet on weight, body composition, serum levels of some hepatokines and adipocytokines, and the profile of EMPs in adults with metabolic syndrome. Moreover, the effects of different diet delivery modes, including “fixed diet plan” and “calorie counting” will also be analyzed. The results of this trial can provide clinical witnesses on the effectiveness of carbohydrate-restricted diets in ameliorating metabolic status and prevent the development of chronic diseases. Trial registration Iranian Registry of Clinical Trials IRCT2016121925267N4. Registered on 26 July 2017

Potency Biomarker Effect of Endothelial Microparticles (EMPs) for Early Prediction of Cardiovascular Risk in Shift Worker Nurses

Objectives. Shift work results in changing worker’s behavior, food, and sleep patterns, which can cause circadian rhythm disturbance, which is a cardiovascular risk. Until now, a biomarker of early prediction of cardiovascular risk on shift workers is still not developed. This study aimed to assess the cardiovascular risk of shift worker nurses by detecting endothelial microparticles (EMPs). Methods. This longitudinal study compared six shift nurses and five non-shift nurses by measuring the EMPs using antigen CD31+ flow cytometry. All met the inclusion criteria consisting of 28 blood samples followed in one week shift. Results. EMPs among non-shift nurses were below 200 μL. However, shift nurses’ EMPs increased above 200 μL with Man-Whitney U p = 0.000 on days 4 and 7 following a one shift per week schedule. Conclusion. There was an increase in shift workers’ endothelial microparticles (EMP) which was a sign of cardio-vascular risk.

Biomarkers of Cardiovascular Toxicity of Benzene Inhalation in Mice

ABSTRACTBenzene is a ubiquitous environmental pollutant. Recent population-based studies suggest that benzene exposure is associated with an increased risk for cardiovascular disease. However, it is unclear whether benzene exposure is sufficient to induce cardiovascular toxicity. We examined the effects of benzene inhalation (50 ppm, 6 h/day, 5 days/week, 6 weeks) or HEPA-filtered air exposure on the biomarkers of cardiovascular toxicity in male C57BL/6J mice. Benzene inhalation significantly increased the biomarkers of endothelial activation and injury including endothelial microparticles, activated endothelial microparticles, endothelial progenitor cell microparticles, lung endothelial microparticles, and activated lung and endothelial microparticles while having no effect on circulating levels of endothelial adhesion molecules, endothelial selectins, and biomarkers of angiogenesis. To understand how benzene may induce endothelial injury, we exposed human aortic endothelial cells to benzene metabolites. Of metabolites tested, trans,trans-mucondialdehyde (10 μM, 18h) was most toxic. It induced caspases-3, −7 and −9 (intrinsic pathway) activation, and enhanced microparticle formation by 2.4-fold. Levels of plateletleukocyte aggregates, platelet macroparticles, and proportion of CD4+ and CD8+ T-cells were also significantly elevated in the blood of the benzene-exposed mice. We also found that benzene exposure increased the transcription of genes associated with endothelial cell and platelet activation in the liver; and induced inflammatory genes and suppressed cytochrome P450s in the lungs and the liver. Together, these data suggest that benzene exposure induces endothelial injury, enhances platelet activation and inflammatory processes; and circulatory levels of endothelial cell and platelet-derived microparticles and platelet-leukocyte aggregates are excellent biomarkers of cardiovascular toxicity of benzene.HighlightsInhaled benzene exposure increases the levels of blood endothelial microparticles.In vitro, benzene metabolite trans, trans-mucondialdehyde induces endothelial cell apoptosis and microparticles formation.Inhaled benzene exposure decreases the levels of hematopoietic progenitor cells in the bone marrow.Inhaled benzene exposure augments the circulating levels of platelet-leukocyte adducts.

Significance of Plasma Circulating Endothelial Microparticles Combined with Von Willebrand Factor in Coronary Injury of Kawasaki Disease

Background:The greatest complication of Kawasaki disease (KD) is coronary artery injury, and the requirement for early diagnosis and treatment is paramount. Thus, markers of vascular endothelial injury are of important clinical significance. Methods:According to our diagnostic criteria, blood samples were collected from 43 patients with KD, who were then divided into coronary artery lesions (CALs) and non-CALs (NCALs) groups according to their Z-score. As the control group, an additional 26 blood samples were collected from healthy children. Flow cytometry (FCM) and enzyme-linked immunosorbent assays (ELISA) were used to detect the expression levels of plasma endothelial microparticles (EMPs) and von Willebrand factor (vWF). Results:The expression levels of plasma CD31+/CD42b-EMPs, CD105+/CD54+EMPs, and vWF were higher in children with KD than those in the control group, and the differences were statistically significant (P<0.05).Also, the expression levels of CD31+/CD42b-EMPs, CD105+/CD54+EMPs and vWF in those in the CALs group at the acute and subacute stages were significantly higher than those in the NCALs group (P<0.05). Furthermore, receiver operating characteristic (ROC) curve analysis revealed that the area under the curve (AUC) of CD31+/CD42b-EMPs combined with vWF was 0.896, which indicates a higher diagnostic value in predicting CALs in children with KD. Conclusions:In our study, expression levels of EMPs and vWFare expected to used for early diagnosis, and which are associated with coronary artery injury in KD.

IL-33 / ST2 Signaling Promotes TF Expression by Regulating NF-κB Activation in Coronary Artery Endothelial Microparticles

IntroductionInterleukin (IL)-33 was previously shown to induce angiogenesis and inflammatory activation of endothelial Microparticles(EMPs). Tissue factor (TF) plays a central role in hemostasis and thrombosis.Material and methodsThe study analyzed the coronary blood of level of CD31+EMPs, TF protein and IL-33 protein in Acute Myocardial Infarction (AMI) and stable coronary artery disease (SCAD) patients. Human coronary artery endothelial cells (HCAECs) were treated with IL-33 to obtain EMPs. The TF activity of EMPs was tested by Thermo Fisher by adding the TF antibody. Furthermore, TF and Tissue Factor Pathway Inhibitor (TFPI) protein were tested by ELISA. Finally, NF-κB inhibitor dimethyl fumarate (DMF) and soluble extracellular domain of ST2 coupled to the Fc fragment of human IgG1 (sST2) were added to HCAECs which were treated with IL-33, and the TF protein level was also tested by ELISA.ResultsThe AMI patients had higher level of CD31+EMPs, TF protein and IL-33 protein than the SCAD patients in coronary artery. In AMI patients (N=27), the IL-33 protein positively correlated with CD31+EMPs (r=0.794, p<0.01). According to the ROC curve analysis, the AUC of CD31+EMPs, TF protein and IL-33 protein were 0.888, 0.962 and 0.778 respectively. In the cell culture, the TF activity and TF protein in EMPs increased gradually with time of intervention by the treatment of IL-33. IL-33 binding to the ST2 receptor promoted TF expression by regulating NF-κB activation in EMPs of HCAECs.ConclusionsActivated endothelial cells and EMPs they released simultaneously express TF, which is a risk factor for cardiovascular disease.