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Atherosclerosis risk factors and macrophage polarization in adipose tissue
Králová, Anna ; Poledne, Rudolf (advisor) ; Vrablík, Michal (referee) ; Zicha, Josef (referee)
Atherosclerosis is a major cause of cardiovascular disease, and the pro-inflammatory state stimulates the process of atherogenesis. Since the 1990s, it has become clear that adipose tissue has not only an endocrine function, but it is also a very important immune organ due to the presence of various immunocytes with high cytokine secretion. Local resident macrophages represent a major component in the development of this inflammation. Due to our inclusion in the unique transplantation program at IKEM, which ensured availability of peroperatively harvested adipose tissue and its subsequent and immediate analysis, we were able to characterize the different subpopulations of monocytes/macrophages and classify macrophages into pro- and anti-inflammatory populations based on the differential expression of surface markers. From subcutaneous (SCAT), perivascular (PVAT), and visceral (VAT) adipose tissue samples from healthy living kidney donors (LKD), we analyzed the macrophage phenotype after obtaining the stromal fraction by flow cytometry. At the beginning of the experiments, we chose the calprotectin molecule as a proinflammatory marker. However, in subsequent experiments described herein, we chose a different route and selected the CD16 receptor to determine the proinflammatory phenotype. This...
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The biological significance of changes in the vascular wall and atherosclerotic plaque for the development of atherosclerotic disease of the coronary arteries
Horváth, Martin ; Hájek, Petr (advisor) ; Poledne, Rudolf (referee) ; Kovárník, Tomáš (referee)
Acute complications of atherosclerosis are typically caused by arterial occlusion due to acute thrombosis forming over a ruptured vulnerable atherosclerotic plaque (VP). Early identification of such VP prior to their rupture could help prevent these events. At the same time, methods for assessing cardiovascular risk are focused on measures at the population level and are not sensitive and specific enough to assess the risk of individual patients. We therefore focused on the possibilities of in vivo detection of VP using biomarkers and invasive imaging methods. First, we focused on the detection of microRNAs (miRs), which have been investigated in recent years as promising biomarkers for a number of diseases, including atherosclerosis. We determined miR levels using quantitative polymerase chain reaction in peripheral venous blood samples from patients in very early phase of ST-elevation myocardial infarction (STEMI), which served as the closest clinical model of acute VP rupture. The results led to the identification of two miRs (miR-331 and miR-151-3p) that were deregulated in STEMI and could be biomarkers of VP. In the next part of the research, we dealt with the possibilities of invasive imaging of VP using intravascular ultrasound and near- infrared spectroscopy (IVUS and NIRS). We confirmed...
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Lipidomic analysis of adipose tissue in patients with ischemic heart disease
Tomášová, Petra ; Vecka, Marek (advisor) ; Poledne, Rudolf (referee) ; Neckář, Jan (referee)
The ever-increasing incidence of coronary artery disease (CAD) in Czech Republic and worldwide is one of the most important health problem today. Obesity and type 2 diabetes mellitus (DM2T) are risk factors for the development of CAD. Lipid metabolism is greatly affected by these diseases and adipose tissue, previously considered a mere energy store, also plays an important role in these changes. Today adipose tissue is discussed also as an endocrine organ, which effect whole body metabolism significantly. Lipidomic analysis is an effective tool for monitoring lipid metabolism. Lipidomics combines the advantages of determining a large number of analytes and a small amount of sample and its rapid preparation. In the first part of study, I optimized the HPLC-MS method of untargeted lipidomic analysis in adipose tissue. Then, this method was used to compare subcutaneous and epicardial adipose tissue. Finally, I evaluated the effect of CAD and DM2T on the lipid profiles of subcutaneous and epicardial adipose tissues. Optimization of the pre-analytical phase enabled the analysis of 206 lipid molecules (in total) in both types of adipose tissue. Compared to subcutaneous adipose tissue, epicardial adipose tissue contained a higher proportion of phosphatidylcholines (e.g. PC(32:0), PC(32:1), PC(36:4)) and...
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The use of PAPP-A/proMBP complex in early diagnosis and prevention of coronary artery disease and in the improvement of therapeutic and preventive care of patients and their families in risk.
Hájek, Petr ; Macek, Milan (advisor) ; Kalousová, Marta (referee) ; Poledne, Rudolf (referee)
Majority of medical decisions are based on results of diagnostic tests that help to differentiate normal from abnormal. The choice of appropriate test and its intepretation are neccessary steps for correct diagnosis and treatment strategy determination. Rapid prove of acute coronary syndrome (ACS) plays a key role in choice of optimal treatment strategy, because timing of intervention directly influences prognosis of the patient. Pregnancy-associated plasma protein-A (PAPP- A) has been studied as a promising marker of ACS. For PAPP-A evaluation in patients with coronary atherosclerosis, we have chosen commercially available system Kryptor that had been verified in prenatal screening of pregnancies in risk. PAPP-A belongs among metalloproteinases. It is important marker of physiological development of placenta and fetus. The only proven physiological role of PAPP- A is the enabling of bioavailability of insulin-like growth factor (IGF). IGF as a growth factor, plays significant role in atherosclerosis development, but also it might contribute to healing processes connected with tissue injury. Nevertheless, PAPP-A role in plaque destabilization has not been proven yet, although it was found in other metaloproteinases. In our pilot study, we confirmed the use of Kryptor system also for patients with coronary...
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The role of intracellular iron in lymfo/monocytes in atherogenesis process
Riško, Peter ; Kraml, Pavel (advisor) ; Poledne, Rudolf (referee) ; Karásek, David (referee)
THE ROLE OF INTRACELLULAR IRON IN LYMFO/MONOCYTES IN ATHEROGENETIC PROCESS MUDr. Peter Riško Background: The increased storage iron level is connected to a higher frequency of cardiovascular events (CVE). Aims and hypothesis: The study investigates the relationship between LIP (labile iron pool) in circulating monocytes and markers of iron metabolism and atherosclerosis (inflammation, oxidative stress, endothelial dysfunction and arterial elasticity) in patients with chronic cardiovascular disease and in healthy volunteers and in long-term blood donors and healthy non-donor volunteers. Methods: A detailed medical history was obtained from the probands and anthropometric measurements (body height, weight and waist, calculation of body mass index and waist-hip ratio) were performed. The following parameters were measured / calculated for all respondents: blood pressure (systolic / diastolic), ankle-brachial index (ABI), pulse pressure and mean arterial pressure, cardio-ankle vascular index (CAVI), augmentation index (AI), pre-ejection time (PEP) and ejection time (ET), sonographic examination of abdomen and carotid arteries were performed. Blood samples were collected in the morning after at least 10 hours of fasting. We performed a basic biochemical examination and measured the parameters of inflammation,...
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Šejda, Tomáš ; Poledne, Rudolf (advisor) ; Filipovský, Jan (referee) ; Češka, Richard (referee)
Based on the results of our study, we concluded that the degree of brachial artery flow endothelium-dependent dilatation is difficult to evaluate using predefined cut-off points as a single-measurement screening test to indicate endothelial dysfunction. On the contrary, the variation of endothelial function after a defined stimulus (dietary, medication) examined at a predefined interval using an identical protocol can help to assess enhanced or reduced efficacy of endothelium-dependent vasodilatation. In the next study, we demonstrated that a high-fat load applied at a single dose does not significantly affect brachial artery vasoreactivity in young, healthy volunteers after four weeks of low- and high-fat diets. In contrast with atherosclerosis, the effect of hypercholestrolemia on adhesive molecule levels has been a source of controversy. We demonstrated that 3-month therapy with fluvastatin does not decrease cICAM-1 levels despite normalization of cholesterol levels. The implication is cholesterol may not induce endothelial activation by the initial upregulation of this adhesive molecule. Endothelial dysfunction can be considered the initial, functionally relevant stage of atherosclerosis, demonstrable still before morphological changes. In patients with advanced obliterative atherosclerosis,...
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Pathophysiology and clinical implications of late coronary thrombosis after implantation of drug eluting stents in patients with manifest atherosclerosis
Jakabčin, Jozef ; Červinka, Pavel (advisor) ; Poledne, Rudolf (referee) ; Mates, Martin (referee)
Pathophysiology and clinical implications of late coronary thrombosis after implantation of drug eluting stents in patients with manifest atherosclerosis Abstract Drug-eluting stents (DES) have markedly reduced restenosis rates compared with bare metal stents (BMS) in controlled randomized trials. Concerns have been raised about a possibly increased incidence of stent thrombosis (ST) after discontinuation of dual antiplatelet treatment compared with BMS. While a restenosis is relatively benign process, this serious complication is an issue because mortality of such event is reaching almost 50%. Among well recognized predictors of late stent thrombosis (delayed endothelisation, renal failure, bifurcation lesions, diabetes, premature antiplatelet therapy discontinuation, slow thrombus disappearance), the stent under expansion and stent deployment technique are also considered to be a contributor for development of late stent thrombosis. There was a paucity of data regarding the IVUS guidance during DES implantation. The aim of this study was to assess the role of IVUS guidance during implantation of DES on longterm outcome in patients with high clinical and angiographic risk profile. Methods: Between January 2004 and December 2005, 2110 patients underwent percutaneous coronary intervention, including 276...
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Factors influencing the metabolism of homocysteine in selected diseases with complex etiopathogenesis
Veselá, Kamila ; Anděl, Michal (advisor) ; Nečas, Emanuel (referee) ; Rušavý, Zdeněk (referee) ; Poledne, Rudolf (referee)
At present, the great medicine to develop molecular genetic techniques, which significantly help to explain the causes of illnesses. In some diseases the genetic basis is well known. These are primarily contingent on monogenic diseases. Unfortunately, many diseases remain the genetic cause is still unknown. The largest group is the so-called complex diseases, where the origin and development of disease involving both genetic and nongenetic factors. These diseases include atherosclerosis, diabetes mellitus, orofacial clefts, preeclampsia, neural tube defects, and many others. Complex diseases are serious health and social problem in developed countries. Clasification of risk factors is the subject of much attention, because knowledge of these factors offer opportunities for effective prevention and treatment. The aim of my work was to obtain new knowledge of factors affecting homocysteine metabolism with regard to the formation of some complex diseases in the Czech population.
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Effect of selected nutrients on skeletal muscle mitochondrial metabolism
Tůmová, Jana ; Anděl, Michal (advisor) ; Poledne, Rudolf (referee) ; Drahota, Zdeněk (referee)
Skeletal muscle plays an important role in the maintenance of whole-body metabolic homeostasis. Metabolic alterations of skeletal muscle contribute to the pathogenesis of a wide range of human diseases, such as obesity, type 2 diabetes and hypertension. Relative excess and suboptimal composition of nutrients negatively affect skeletal muscle metabolism and a better understanding of mechanisms involved in these changes is of central importance. The aim of the work presented in this thesis was to explore cell viability and mitochondrial respiratory parameters following experimentally induced changes in the availability or composition of selected nutrients (fatty acids and glutamine). We attempted to elucidate the mechanisms responsible for the observed changes, such as mitochondrial DNA (mtDNA) damage, or nuclear receptors activation. The studies were performed in vitro on skeletal muscle cell culture models. In addition, we examined mitochondrial function and fat accumulation in skeletal muscle of vegans, i.e. subjects consuming a strict plant-based diet. Using C2C12 skeletal muscle cells we studied the effects of free fatty acids (FFA). We found that relatively low doses of saturated palmitic acid increased hydrogen peroxide production and induced mtDNA damage, mitochondrial respiratory dysfunction...
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Iron metabolism, oxidative stress and progression of atherosclerosis
Syrovátka, Petr ; Kraml, Pavel (advisor) ; Poledne, Rudolf (referee) ; Vokurka, Martin (referee)
Ferritin, oxidative stress and insulin resistance Aim: The aim of our cross-sectional study was to assess the relationships between body iron stores, oxidative stress and impaired insulin sensitivity in a cohort of healthy men in primary prevention of cardiovascular disease. Methods: We examined 151 volunteers, aged 35- 60 years. Anthropometric parameters, markers of metabolic syndrome, insulin resistance, inflammatory markers, parameters of oxidative stress and parameters of endothelial dysfunction were measured. Results: Ferritin correlated positively with waist circumference, body mass index, impaired insulin sensitivity, plasma triglycerides and inversely with high-density lipoprotein cholesterol. We observed positive correlations between ferritin, oxidized low-density lipoprotein and advanced oxidation protein products after adjustment for age, waist circumference, body mass index and measured inflammatory markers (high-sensitivity C-reactive protein, fibrinogen, interleukin-6 and tumor necrosis factor-α). In a stepwise multiple regression analysis, triglycerides, waist circumference and elevated transaminases were independent determinants of the serum ferritin level. Conclusions: Our results provide evidence for a relationship between plasma ferritin and oxidative modification of lipids as...
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