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Effect of atorvastatin on endoglin expression and function in hyperlipidemia-induced endothelial dysfunction
Holadová, Adéla ; Tripská, Katarína (advisor) ; Fikrová, Petra (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Title of Diploma Thesis: Effect of atorvastatin on endoglin expression and function in hyperlipidemia-induced endothelial dysfunction Author: Adéla Holadová Supervisor of Diploma Thesis: PharmDr. Katarína Tripská, Ph.D. Aim: The aim of this thesis was to determine how atorvastatin affects the expression and function of endoglin and related biomarkers of inflammation/endothelial dysfunction at different concentrations and times in human endothelial cells from the aorta, in which hyperlipidemia was induced by using palmitic acid. Methods: In the diploma thesis, we worked with human aortic endothelial cells. We premedicated them with atorvastatin/palmitic acid at different times and concentrations. Using flow cytometry, we measured the protein expression of endoglin. Furthermore, we used the qRT-PCR method to determine the level of mRNA expression of endoglin, its transcription factors and biomarkers of inflammation. Using the ELISA method, we detected the levels of soluble endoglin. Results: We demonstrated that atorvastatin at a concentration of 5μM after 16 hours of premedication significantly increased the mRNA expression of endothelial nitric oxide synthase, and conversely decreased the...
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Effect of glipizide on the expression and function of endoglin and related biomarkers of endothelial dysfunction in diabetes mellitus type II
Vochozková, Iva ; Tripská, Katarína (advisor) ; Urbánková Rathouská, Jana (referee)
8 ABSTRACT Charles University, Faculty of Pharmacy in Hradec Kralové Department of Biological and Medical Sciences Title of Diploma Thesis: Effect of glipizide on the expression and function of endoglin and related biomarkers of endothelial dysfunction in type II diabetes mellitus Author: Iva Vochozková Supervisor of Diploma Thesis: PharmDr. Katarína Tripská, Ph.D. Aim: The aim of this thesis was to determine how glipizide affects the expression and function of endoglin, its transcription factors and related biomarkers of endothelial dysfunction, soluble endoglin levels and monocyte adhesion to human diabetic coronary artery endothelial cells. Methods: In this thesis we worked with human diabetic coronary artery endothelial cells. These were premedicated with glipizide at various times and concentrations. We measured protein expression of endoglin and biomarkers of endothelial dysfunction using flow cytometry. We also measured monocyte adhesion to endothelial cells using flow cytometry. We detected soluble endoglin levels by ELISA and measured mRNA expression of endoglin, its transcription factors and markers of inflammation by PCR. Results: We demonstrated that premedication with 200µM concentration of glipizide led to a decrease in endoglin protein expression after 24 and 48 hours. The protein expression...
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Evaluation of endoglin and P-selectin expression and co-expression in aortas of apoE-deficient mice
Brlicová, Monika ; Rathouská, Jana (advisor) ; Kovařík, Miroslav (referee)
Charles University in Prague Faculty of Pharmacy in HradecKrlov Department of Biological and Medical Sciences Evaluation of endoglin and P-selectin expression and co-expression in aortas of apoE-deficient mice Diploma thesis MonikaBrlicov Supervisor: Mgr.JanaRathousk Background: We observed the expression and the reciprocal co-expression of endoglin (receptor III for TGF- cytokine) and P-selectin (adhesion molecule and marker of endothelial dysfunction) in ascending aortas of apoE-deficient mice which were fed by standard diet for rodents and Western type diet (high-cholesterol diet) for achieving of different phases of the atherosclerotic process. The changes of total cholesterol levels in mice after administration of both types of diets were also evaluated. Methods: The modified strain C57BL/6J of mice with a deficiency of apolipoprotein E, which is prone to aterogenesis was used for this diploma thesis. Mice were divided into three groups. The first group was fed by standard diet (so-called "chow" diet) for a period of two months and the second two groups were fed by Western type diet for a period of two and four months. The levels of total cholesterol in the blood were biochemically determinated and then we statistically evaluated this levels in all groups. Immunohistochemical...
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Cardiovascular Risks in Chronic Airway Disease in Childhood
Kreslová, Marcela ; Sýkora, Josef (advisor) ; Fila, Libor (referee) ; Kopřiva, František (referee)
1 Cardiovascular risks in chronic airway disease in childhood The aim of this thesis was to evaluate cardiovascular risk by using a combined diagnostic approach by measuring RHI and specific biochemical markers in patients with chronic respiratory disease, where we could assume a possible risk of CVD. A total of 119 probands were examined, including 22 patients with cystic fibrosis (CF) and 52 asthma patients. We evaluated RHI using a new plethysmographic method that has a number of advantages over the ultrasonographic methods used in other studies, including non-invasiveness, high sensitivity, low biological variability and objectivity due to automatic processing. Of the biochemical parameters, we measured 4 biomarkers in relation to endothelial dysfunction (ED): hsCRP, ADMA, E-selectin, and VCAM-1. We compared RHI and biomarkers in CF and asthma patients with healthy controls and sought mutual correlations. We did not prove a statistically significant difference in RHI between the test groups with CF children but we confirmed the decreasing trend of RHI since adolescence and significantly lower RHI values in CF adults, confirming the progressive development of atherogenesis and worsening of ED with age. Biochemical parameters showed significantly higher levels of hsCRP, sVCAM-1 and E-selectin in CF...
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Molecular basis of endothelial sysfunction: endothelial nitric oxide synthase and heme oxygenase 1 genetic variations
Král, Aleš ; Martásek, Pavel (advisor) ; Baxová, Alice (referee) ; Schneider, Bohdan (referee)
Endothelial dysfunction is a pathologic state characterized by an altered equilibrium among vasodilatory and antithrombotic mediators and vasoconstrictive and prothrombotic mediators produced by the vascular endothelium. Multiple factors induce impaired production or increased consumption nitric oxide (NO), the key mediator of vascular homeostasis, produced by the nitric oxide synthase enzymes (NOS). Endothelial dysfunction represents one of the initial steps in the development of atherosclerosis, a chronic inflammatory disease of the vascular wall. The inducible enzyme heme oxygenase 1 (HO-1) represents one of the main cellular defense mechanisms against increased oxidative stress and decreased NO bioavailability accompanying endothelial dysfunction and atherosclerosis. We studied the genetic determinants of endothelial dysfunction and atherosclerosis by evaluating the association of the G894T endothelial NOS (eNOS) polymorphism and the HO-1 (GT)n promoter polymorphism with coronary artery atherosclerosis severity and risk profile and their evolution during hypolipidaemic treatment. In addition, we searched for genetic variations in exons 25 and 26 of eNOS gene, encoding the C-terminal part of the protein, deemed crucial for proper enzyme function and the 3'- untranslated region crucial for eNOS...
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Relationship of oxidative stress to parameters of diabetes control in development of vascular complications.
Pelcl, Tomáš ; Prázný, Martin (advisor) ; Rušavý, Zdeněk (referee) ; Šmahelová, Alena (referee)
The aim of this thesis is to contribute to the clarification of the pathogenesis of chronic complications of diabetes mellitus. The main goal of the research was glycaemic variability, its contribution to the activation of oxidative stress and its possible role in the process of advanced glycation, all beyond the scope of persistent hyperglycaemia itself. Another aim of the work is to contribute to the clarification of a possible relationship between glycaemic variability and vascular complications of diabetes. We were the first to describe the association between the concentrations of reactive aldehydes formed during lipid peroxidation and disorders of skin microvascular reactivity in patients with type 1 diabetes (DM1). Elevated markers of oxidative stress were found in this group, furthermore during the 3 years of follow-up higher plasma antioxidant activity was observed. These findings were not dependent of the method of glucose monitoring and glucose variability, which was lower in a subgroup of patients using real-time continuous glucose monitoring (rt-CGM), compared to a subgroup using conventional glucometers. However, it is clear, that hyperglycaemia alone induces increased oxidative stress in patients with diabetes. Simultaneously we observed the opposite process of oxidative stress...
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Searching for and Evaluating the Severity of Endothelial Dysfunction in Children with Chronic Autoimmune Disease
Sýkorová, Aneta ; Jehlička, Petr (advisor) ; Urbanová, Zuzana (referee) ; Klásková, Eva (referee)
We aimed to evaluate the endothelial function by combining RHI measurements and specific biochemical markers in the children with possible risk of premature manifestation of atherosclerosis and in the control group of healthy children. In all, 124 children (of which 106 patients divided into five groups according to diagnosis - type 1 diabetes mellitus, Crohn's disease, cystic fibrosis, familial hypercholesterolemia and acute lymphoblastic leukemia and 18 healthy controls) were enrolled in the study. During the study, we measured RHI using a new plethysmographic method and further evaluated biochemical markers of endothelial dysfunction (ADMA, E-selectin, hsCRP and VCAM) and lipidogram in individual groups of children. The primary objective of our study was the determination of RHI and biochemical parameters in healthy subjects and in selected risk groups of children (type 1 diabetes mellitus, Crohn's disease, cystic fibrosis, familial hypercholesterolemia and children after successful treatment of acute lymphoblastic leukemia). At the same time, we compared patients from individual groups with the control group. We found significantly elevated RHI values in groups of children with type 1 diabetes, Crohn's disease, cystic fibrosis, and children after successful treatment of acute lymphoblastic leukemia....
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Evaluation of endoglin and P-selectin expression and co-expression in aortas of apoE-deficient mice
Brlicová, Monika ; Rathouská, Jana (advisor) ; Kovařík, Miroslav (referee)
Charles University in Prague Faculty of Pharmacy in HradecKrlov Department of Biological and Medical Sciences Evaluation of endoglin and P-selectin expression and co-expression in aortas of apoE-deficient mice Diploma thesis MonikaBrlicov Supervisor: Mgr.JanaRathousk Background: We observed the expression and the reciprocal co-expression of endoglin (receptor III for TGF- cytokine) and P-selectin (adhesion molecule and marker of endothelial dysfunction) in ascending aortas of apoE-deficient mice which were fed by standard diet for rodents and Western type diet (high-cholesterol diet) for achieving of different phases of the atherosclerotic process. The changes of total cholesterol levels in mice after administration of both types of diets were also evaluated. Methods: The modified strain C57BL/6J of mice with a deficiency of apolipoprotein E, which is prone to aterogenesis was used for this diploma thesis. Mice were divided into three groups. The first group was fed by standard diet (so-called "chow" diet) for a period of two months and the second two groups were fed by Western type diet for a period of two and four months. The levels of total cholesterol in the blood were biochemically determinated and then we statistically evaluated this levels in all groups. Immunohistochemical...
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Molecular basis of endothelial sysfunction: endothelial nitric oxide synthase and heme oxygenase 1 genetic variations
Král, Aleš ; Martásek, Pavel (advisor) ; Baxová, Alice (referee) ; Schneider, Bohdan (referee)
Endothelial dysfunction is a pathologic state characterized by an altered equilibrium among vasodilatory and antithrombotic mediators and vasoconstrictive and prothrombotic mediators produced by the vascular endothelium. Multiple factors induce impaired production or increased consumption nitric oxide (NO), the key mediator of vascular homeostasis, produced by the nitric oxide synthase enzymes (NOS). Endothelial dysfunction represents one of the initial steps in the development of atherosclerosis, a chronic inflammatory disease of the vascular wall. The inducible enzyme heme oxygenase 1 (HO-1) represents one of the main cellular defense mechanisms against increased oxidative stress and decreased NO bioavailability accompanying endothelial dysfunction and atherosclerosis. We studied the genetic determinants of endothelial dysfunction and atherosclerosis by evaluating the association of the G894T endothelial NOS (eNOS) polymorphism and the HO-1 (GT)n promoter polymorphism with coronary artery atherosclerosis severity and risk profile and their evolution during hypolipidaemic treatment. In addition, we searched for genetic variations in exons 25 and 26 of eNOS gene, encoding the C-terminal part of the protein, deemed crucial for proper enzyme function and the 3'- untranslated region crucial for eNOS...
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Aldosterone synthase in arterial hypertension and possible influence of its genenetic polymorphism on left ventricular hypertrophy
Heller, Samuel ; Horký, Karel (advisor) ; Málková, Jana (referee) ; Widimský, Jiří (referee)
Part I. The aldosterone synthase gene (CYP11B2) polymorphism T-344C in blood pressure and left ventricular hypertrophy. BACKGROUND: Aldosterone is a key cardovascular hormone, it significantly influences volume, pressure and electrolyte balance. Aldosterone plays an important role in development of left ventricular (LV) hypertrophy and myocardial fibrosis. The aldosterone synthase gene (CYP11B2) is an important candidate gene region in essential hypertension. DESIGN AND METHODS: We assessed the influence of the T-344C polymorphism of aldosterone synthase - the rate-limiting enzyme in aldosterone biosynthesis - on the structure of the left ventricle in young normotensive men. The population included 113 normotensive mid-European Caucasian men aged 18-40 years (mean 27 +/- 5 years). We also studied the association of -344T/C polymorphism of the CYP11B2 gene with the presence and severity of hypertension in 369 individuals, of whom 213 were hypertensive patients (139 controlled hypertensive, 74 resistant hypertensive) and 156 were healthy normotensive subjects. The genotype was assessed using polymerase chain reaction with subsequent cleavage with restriction enzyme HAEIII (restriction fragment length polymorphism method) and visualization with ethidium bromide. Plasma renin activity (PRA) and plasma...
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