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Tissue and soluble endoglin relation to the endothelial dysfunction and possible treatment
Blažíčková, Kateřina ; Nachtigal, Petr (advisor) ; Blaha, Vladimír (referee) ; Mičuda, Stanislav (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Candidate: Msc. Katerina Blazickova Supervisor: Assoc. Prof. PharmDr. Petr Nachtigal, Ph.D. Title of Doctoral Thesis: Tissue and soluble endoglin relation to the endothelial dysfunction and possible treatment Atherosclerosis is a complex inflammatory disease and it represents a major source of morbidity and mortality in the world as a part of cardiovascular diseases. Endothelial dysfunction represents the first step in the development of atherosclerosis and it is characterized by disruption of endothelial homeostasis. Statins are drugs of choice in the treatment of atherosclerosis and they can decrease LDL cholesterol levels and positively affect levels of HDL cholesterol. Statins, however have a wide range of other effects, which are referred to non-lipid or pleotropic effects. The most important non-lipid effects of statins are lipid- independent modulation of endothelial function, antioxidant, anti-inflammatory, antithrombogenic and antiproliferative effects. The most widely used animal model for the study of atherosclerosis is the mouse model. Thanks to a high fat diet and genetic modification, we are able to modify lipid profile in mice and induce atherosclerotic changes even at...
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Early changes in tissue endoglin and VCAM-1 expression in mice aorta after administration of high fat diet
Swierczková, Iva ; Nachtigal, Petr (advisor) ; Herink, Josef (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Consultant: doc. PharmDr. Petr Nachtigal, Ph.D. Candidate: Bc. Iva Swierczková Title of Thesis: Early changes in tissue endoglin and VCAM-1 expression in mice aorta after administration of high fat diet Background: The aim of the thesis was to observe changes in tissue endoglin and VCAM-1 expression in mice aorta after administration of chow and high-fat diet using immunohistochemical methods. Finally, these two groups were compared. Method: Two-month-old female C57BL/6J mice were used for the experiment. First half, which served as a control group, was fed with the chow diet. The second half was fed with the high fat diet, which contains 1.25 % of cholesterol and 40 % of fat, for the period of 3 months. Immunohistochemical analysis was performed using the indirect three-step avidin-biotin complex method. Results: The expression of endoglin was detetced on the vascular endothelium but there were no evident differences in the staining intensity between the control group and the experimental group. We also observed VCAM-1 expression in the vascular endothelium. In the control group, expression appeared to be stronger than the experimental group, however these differences were not that significant....
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Monitoring the effect of statins and soluble endoglin on markers of endothelial dysfunction in selected cell line and culture.
Vařejčková, Michala ; Nachtigal, Petr (advisor) ; Štěrba, Martin (referee) ; Mladěnka, Přemysl (referee)
Charles University Faculty of Pharmacy in Hradec Kralove Department of Biological and Medical Sciences Candidate: Mgr. Michala Vařejčková Supervisor: Prof. PharmDr. Petr Nachtigal, Ph.D. Title of Doctoral Thesis: Monitoring the effect of statins and soluble endoglin on markers of endothelial dysfunction in selected cell line and culture. This doctoral thesis studied the effect of soluble endoglin on endothelial dysfunction markers in vitro in primary cell culture HUVEC and cell line HEK293T. Some experiments included in this thesis were made in cooperation in the laboratory of prof. Carmelo Bernabeu in the research center Centro de Investigaciones Biológicas (CIB) in Madrid. Endothelial dysfunction is damage of endothelium when the balance between vasoconstrictor and vasodilatory mechanisms is disrupted, it's leading to platelet aggregation, leukocyte adhesion and increased permeability of the endothelium. All these events represent the first step in the development of atherosclerosis and they are part of other cardiovascular diseases such as preeclampsia, type 2 diabetes mellitus, familial hypercholesterolemia. An important role in these diseases play endoglin, eNOS and soluble endoglin (sEng), which is cleaved by MMP- 14 from the membrane endoglin and released into the circulation. It was found...
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Early changes in tissue endoglin and VCAM-1 expression in mice aorta after administration of high fat diet
Swierczková, Iva ; Nachtigal, Petr (advisor) ; Herink, Josef (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Consultant: doc. PharmDr. Petr Nachtigal, Ph.D. Candidate: Bc. Iva Swierczková Title of Thesis: Early changes in tissue endoglin and VCAM-1 expression in mice aorta after administration of high fat diet Background: The aim of the thesis was to observe changes in tissue endoglin and VCAM-1 expression in mice aorta after administration of chow and high-fat diet using immunohistochemical methods. Finally, these two groups were compared. Method: Two-month-old female C57BL/6J mice were used for the experiment. First half, which served as a control group, was fed with the chow diet. The second half was fed with the high fat diet, which contains 1.25 % of cholesterol and 40 % of fat, for the period of 3 months. Immunohistochemical analysis was performed using the indirect three-step avidin-biotin complex method. Results: The expression of endoglin was detetced on the vascular endothelium but there were no evident differences in the staining intensity between the control group and the experimental group. We also observed VCAM-1 expression in the vascular endothelium. In the control group, expression appeared to be stronger than the experimental group, however these differences were not that significant....
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The importance of membrane and soluble endoglin in atherosclerosis.
Mphetang, Constance Bontle ; Nachtigal, Petr (advisor) ; Najmanová, Iveta (referee)
This thesis is about the importance of membrane and soluble endoglin (sEng) in atherosclerosis, both the development and the progression of the state. Atherosclerosis poses a major risk factor for heart diseases and stroke, which renders it a serious health issue. The role of endoglin in atherosclerosis is yet to be defined and this will be discussed below. Endoglin expression was discovered in atherosclerotic vessels predominantly in the endothelial and smooth muscle cells both in experimental animals and humans. This discovery suggests that endoglin plays a role in atherogenesis and stabilization of atherosclerotic plaques. In addition to atherosclerosis, high levels of soluble endoglin were also discovered in cases where hypercholesterolemia and myocardial infarction were observed and were also related to TGF-β signalling inhibition in the walls of the affected vessels. Additionally, statins reasonably lowered soluble endoglin levels and facilitated its expression in the aorta of mice, resulting in a decrease in the atherosclerotic effects and complications. Furthermore, patients with familial hypercholesterolemia underwent extracorporeal eliminations which helped significantly reduce the levels of soluble endoglin. In conclusion, knowing the levels of soluble endoglin and what that indicates in...
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Western blot analysis of selected inflammatory markers in aorta of apoE/LDLR deficient mice
Krejcarová, Jana ; Rathouská, Jana (advisor) ; Nachtigal, Petr (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Title of Diploma Thesis: Western blot analysis of selected inflammatory markers in aorta of apoE/LDLR deficient mice Author: Jana Krejcarová Supervisor: PharmDr. Jana Rathouská, Ph.D. Background: This thesis was focused on the detection and subsequent evaluation of the expression of adhesion molecules ICAM-1, VCAM-1 and COX-2 and P-selectin in mouse aortas. Control and monitored group of mice differed in the genetic material, where monitored group of mice had unfunctional genes for apolipoprotein E and LDL receptor (apoE−/− /LDLR−/− ). Methods: For the study, two months old male mice of the C57BL/6J strain were used. Males were divided into two groups - control and monitored with unfunctional receptor for low- density lipoprotein and for apolipoprotein E. Both groups were fed by a standard diet for rodents. For detection and subsequent quantification of the expression of the markers VCAM-1, ICAM-1, COX-2 and P-selectin, Western blot analysis was used. Levels of soluble ICAM-1 and P-selectin in plasma were assessed by Luminex®. Results: Western blot analysis showed no significant difference in the expression of VCAM-1, ICAM-1, COX-2 and P-selectin between both groups of mice. Using the...
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Early changes in eNOS and ICAM-1 expression in mice aorta after administration of high fat diet.
Stehlíková, Tereza ; Nachtigal, Petr (advisor) ; Blažíčková, Kateřina (referee)
Charles University Faculty of Pharmacy in Hradec Kralove Department of Biological and Medical Sciences Consultant: Associate Professor, Petr Nachtigal, Ph.D. Candidate: Tereza Stehlíková Title of Thesis: Early changes in eNOS and ICAM-1 expression in mice aorta after administration of high fat diet Background: The aim of this thesis was to observe changes in eNOS a ICAM-1 expression after administration of high fat diet in comparison with the control group, which was fed with the chow diet. Method: C57BL/6J female mice were used for the experiment. 6 mice were fed with the high fat diet (experimental group), which contains 1,25 % of cholesterol and 40 % of fat, for the period of 3 months. Six mice (representing control group) were fed with the chow diet the whole time. Immunohistochemical analysis was performed by means of avidin-biotin complex method. Results: The expression of eNOS was visible on the vascular endothelium but there were no differences between the control group and the experimental group. Similarly, ICAM-1 expression was detected only in endothelium in aorta but no differences in the expression between control and experimental group were visible. Conclusions: Administration of high fat diet with 1,25 % of cholesterol and 40 % of fat for 3 months had no effect on the expression of...
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Anti-inflammatory effects of ursodeoxycholyl lysophosphatidylethanolamide on THP-1 human macrophages via Toll-like receptor 4
Horvátová, Alžbeta ; Pávek, Petr (advisor) ; Nachtigal, Petr (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Alžbeta Horvátová Supervisor: prof. PharmDr. Petr Pávek PhD. Title of diploma thesis: Anti-inflammatory effects of ursodeoxycholyl lysophosphatidylethanolamide on THP-1 human macrophages via Toll-like receptor 4 Nonalcoholic steatohepatitis (NASH) became the most common liver disease in developed countries. It is well-known that the level of protectant phosphatidylcholine (PC) is decreased in NASH. The bile acid-phospholipid conjugate ursodeoxycholyl lysophosphatidylethanolamide (UDCA-LPE) was designed in order to specifically deliver PC to hepatocytes. However, previous studies have proved that UDCA-LPE possesses its proper hepatoprotectant capacity and exhibits anti-apoptotic, anti-inflammatory, anti-fibrotic properties and also improved steatosis and hyperlipidaemia in various models in vivo. These effects may be mediated secondary through modulation of immune system. Therefore, in order to dissect if UDCA-LPE directly influences immune cells in vitro, release of pro-inflammatory cytokines TNFα, IL-6 and IL-1β in LPS-induced THP-1-derived human macrophages was measured by ELISA. Moreover, effects of UDCA-LPE on MAPK signalling pathways and nuclear translocation of NFκB were...
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Tissue and soluble endoglin relation to the endothelial dysfunction and possible treatment
Blažíčková, Kateřina ; Nachtigal, Petr (advisor) ; Blaha, Vladimír (referee) ; Mičuda, Stanislav (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Candidate: Msc. Katerina Blazickova Supervisor: Assoc. Prof. PharmDr. Petr Nachtigal, Ph.D. Title of Doctoral Thesis: Tissue and soluble endoglin relation to the endothelial dysfunction and possible treatment Atherosclerosis is a complex inflammatory disease and it represents a major source of morbidity and mortality in the world as a part of cardiovascular diseases. Endothelial dysfunction represents the first step in the development of atherosclerosis and it is characterized by disruption of endothelial homeostasis. Statins are drugs of choice in the treatment of atherosclerosis and they can decrease LDL cholesterol levels and positively affect levels of HDL cholesterol. Statins, however have a wide range of other effects, which are referred to non-lipid or pleotropic effects. The most important non-lipid effects of statins are lipid- independent modulation of endothelial function, antioxidant, anti-inflammatory, antithrombogenic and antiproliferative effects. The most widely used animal model for the study of atherosclerosis is the mouse model. Thanks to a high fat diet and genetic modification, we are able to modify lipid profile in mice and induce atherosclerotic changes even at...
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The role of human papilloma viruses in ethiopathogenesis malignant tumors in sinonasal area
Sieglová, Kateřina ; Nachtigal, Petr (advisor) ; Petera, Jiří (referee)
The aim of this thesis is the comparison of immunohistochemical and molecular biological methods for the detection of human papillomavirus (HPV) in tumour tissue and their subsequent use in the study of HPV role in the etiopathogenesis of malignant tumors of sinonasal region. Several methods for the purpose of HPV detection were applied: in situ hybridization, immunohistochemical detection of expression of p16INK4a protein, classic polymerase chain reaction combined with two different enzyme immunoassay detection methods, and reverse hybridization, followed by polymerase chain reaction with real time reverse transcription. All methods (p16 DNA in situ hybridization, RNA in situ hybridization, quantitative polymerase chain reaction with reverse transcription - RT-qPCR mRNA) used for the detection of HPV performed very well when compared, showing concordance with statistical significance p ˂ 0,0001. Detection of p16 vs. DNA polymerase chain reaction (PCR) had correlation coefficient p = 0.004 in the whole group, and p = 0.002 in the subgroup of squamous cell carcinoma (SCC). Correlation coefficient between detection of p16 and overall HPV status was 0,659 for entire set and 0,864 for set of SCC, indicating significant linear dependence. Immunohistochemical detection of p16 in combination with the...
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