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Endothelial dysfunction markers and soluble endoglin
Pešlová, Lenka ; Nachtigal, Petr (advisor) ; Němečková, Ivana (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Kralove Department of Biological and Medical Sciences Consultant: Doc. PharmDr. Petr Nachtigal, PhD. Student: Lenka Pešlová Title of Thesis: Endothelial dysfunction markers and soluble endoglin Background: To analyze possible changes in histological structure in myocardium of mice with low or high levels of soluble endoglin. We focused on description of general histology, observation of ligament changes and possible development of inflammatory reactions using immunohistochemical analysis. Method: In the study we used genetically modified female mice, strain CBAxC57BL/6J, with low or high levels of soluble endoglin (Sol-Eng+). Six-month- old female mice were fed for three months using a high fat rodent diet containing 1,25% of cholesterol and 40% of fat. Then their myocardia were resected. For identification of endoglin expression we used immunohistochemical analysis called En Vision-CD3-T-lymfocyty (rabbit anti mouse). In histological analysis we used hematoxylin-eosin and green trichrome staining. Results: Histological analysis did not reveal any pathological accumulation of collagen in any of animal myocardium we focused on. There were any changes in staining neither of nuclei or cytoplasm, nor any infiltration of leucocytes....
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Immunohistochemical detection of macrophages in atherosclerotic lesions brachiocephalic artery I
Škodová, Sabina ; Nachtigal, Petr (advisor) ; Němečková, Ivana (referee)
Sabina Škodová Immunohistochemical detection of macrophages in atherosclerotic lesions brachiocephalic artery. Bachelor thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Medical Laboratory Technician Background: The aim of this bachelor thesis was to set imunohistochemic methods for detection of the macrophages in brachiocephalic artery apoE/LDLR receptor deficient mice. This thesis also deals with quantification of expression macrophages in group of mice which were fed with chow diet and in group of mice which were fed with low carbohydrate high protein diet. Methods: For this study were used apoE/LDLR deficient female mice which were separated into two groups (in each group were 6 mice). The first group was fed with chow diet (control group) and the second one was fed with low carbohydrate high protein diet (LCHP). This diet was administered to the mice for 8 weeks. After that the imunohistochemical analysis was performed on the cuts of brachiocephalic artery which uses Immpress method with diaminobenzidin (DAB) for better visibility of macrophages. Afterwards was performed biochemical and stereological analysis. Results: Biochemical analysis showed increased levels of total cholesterol in mice with LCHP diet. Immunohistochemical analysis showed presence of macrophages in...
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The study oxidative stress markers in animal model of chronic colitis in large intestine.
Hocková, Erika ; Nachtigal, Petr (advisor) ; Němečková, Ivana (referee)
Crohn's disease and ulcerative colitis are both entities of inflammatory bowel disease (IBD). The nuclear transcription factor kappaB (NFκB) family consists of five different members, which are namely p65 (RelA), c-Rel, RelB, p50 and p52. NFκB has ability to promote the expression of various proinflammatory genes, to influence the course of mucosal inflammation and to mediate different cell-type specific effects. Bilirubin is an endogenous substance with anti-oxidative and anti-inflammatory effects, which influences the process of inflammation and has protective effect. This diploma thesis was aimed at conduction of methods of NFκB detection on paraffine tissue slices and observation of NFκB expression in large intestine walls of normobilirubin and hyperbilirubin rats (Gunn's variety) after chronic imunohistochemically- induced inflammation. Eight Gunn's variety rats (G+ ) and eight Wistar's variety rats (K+ ) were used in this study, all after long time administration of dextrane-sulfate (DSS). For seven following days, DSS (in concentration of 2%w/v) was added into drinkable water for these rats, followed by 14 days of clear drinkable water (without DSS) administered to rats. This three-weeks long cycle was repeated twice (3 cycles in total were completed). Six rats of both varieties (6 G+ and 6...
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The effect of atorvastatin on the expression of markers of the cascade transforming growth factor beta in experimental atherogenesis.
Frnochová, Lenka ; Nachtigal, Petr (advisor) ; Němečková, Ivana (referee)
Lenka Frnochová The effect of atorvastatin on the expression of markers of the cascade transforming growth factor beta in experimental atherogenesis. Diploma thesis Charles Univerzity in Prague, Faculty of Pharmacy in Hradec Králové Pharmacy Background: The aim of this diploma thesis was to observed the expression of ALK-1 and VEGF in atherosclerotic plaques in ApoE and LDL receptor- deficient mice with regard to the use of atorvastatin. Methods: We used for research female C57BL/6J mice with double deficiency of apolipoprotein E and LDL receptor. Mice were 8 weeks old. Mice were divided into two groups: atorvastatin and control group. Both groups were fed cholesterol diet for 8 weeks and in atorvastatin group was also fed with atorvastatin. Then biochemical analysis of blood was performed. For detection of lipids in atherosclerotic lesions we used histological staining by oil red. Immunohistochemical analysis was performed in 1cm aortic sinus and part of aortic arch. The method used to detect the expression of ALK-1 and VEGF was the avidin-biotin method with detection using DAB. Results: Biochemical analysis showed that atorvastatin reduced levels of total cholesterol. After the staining by oil red atherosclerotic lesions were seen in both groups. Atorvastatin reduced the size of atherosclerotic...
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Sunitinib Effects on the Expression of ICAM-1 in Normotensive and Hypertensive Rats
Frnochová, Lenka ; Nachtigal, Petr (advisor) ; Němečková, Ivana (referee)
Mgr. Lenka Frnochová Sunitinib effects on the expression of ICAM-1 in normotensive and hypertensive rats Thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Pharmacy Background: We quantified the expression of ICAM-1 in aorta of normotensive Wistar Kyoto rats and spontaneously hypertensive rats during use of sunitinib. For quantification we applied stereological and immunohistochemical methods. Methods: We used inbred male of Wistar Kyoto rats and spontaneously hypertensive rats for our research. Rats of both strains were divided into two groups - a sunitinib group and a control group. The experimental group was receiving drinking water with sunitinib of 10 mg/kg per day and rats in the control group was receiving only drinking water. Spontaneously hypertensive rats were receiving sunitinib for 8 weeks and after 5 days of regeneration they got sunitinib into the water back for another 8 weeks. Wistar Kyoto rats were receiving sunitinib in water for 8 weeks, but after 5 days of regeneration, these rats were receiving sunitinib only for 2 weeks due to toxicity and weight loss. Immunohistochemical analysis was applied in rat aorta. For detection of the ICAM-1 expression was used En Vision method with DAB detection. Results: Immunohistochemical staining showed that expression of...
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Sunitinib effects on endothelial expression of hemeoxygenase-I in normotensive rats
Svobodová, Barbora ; Nachtigal, Petr (advisor) ; Němečková, Ivana (referee)
Barbora Svobodová Sunitinib effects on endothelial expression of hemeoxygenase-I in normotensive rats. Bachelor thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Medical Laboratory Technician Background: The aim of this bachelor thesis was to define and describe the expression of hemeoxygenase-1(HO-1) in the aortas of normotensive Wistar Kyoto (WKY) rats and evaluate possible effects of sunitinib treatment by means of immunohistochemical and stereological analysis. Methods: Two groups of WKY rats were used for the experiment (6 animals in each of them). Sunitinib was administered to the first group. The other one (the control one) received water only. Application of sunitinib proceeded by following scheme: 8 weeks of using/ 5 weeks rest period/ 2 weeks of using. After that we performed the immunohistochemical analysis on 60 preparations of rat's aortas which observed the expression of HO-1 by methodology En Vision with DAB detection. Then the stereological evaluation was performed. Results: The immunohistochemical analysis showed the expression of HO-1 in the endothelium only and it was noticed just in several animals especially from the group with sunitinib treatment. Conclusion: The immunohistochemical and stereological analysis demonstrated a significant increase of the...
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Molecular genetics of celiac disease
Němečková, Iva ; Daňková, Pavlína (advisor) ; Tučková, Ludmila (referee)
Celiac disease is an organ-specific autoimmune disease that arises as a consequence of hypersensitivity to the grain gluten in genetically susceptible individuals. Genetic predisposition are HLA-DQ2 and HLA-DQ8 genes, which are necessary but not sufficient for the emergence of celiac disease; it is involved in approximately 40% of the inheritance. In the course of the time, other genes that might contribute to the pathogenesis of celiac disease are being discovered. Among these so-called candidate genes, which are sought on the basis of known knowledge of molecular mechanisms of innate and adaptive immune responses, are for example: MIC, TNF, CTLA-4, CD28, ICOS, MYO9B, MMP, TLR and PTPN22. Immune response triggered by gluten peptide penetration into the lamina propria leads to mucosal damage. Different gluten peptides are involved in the pathology of celiac disease in different ways, some peptides trigger an adaptive immune response, while others, such as peptide p31- 43, triggers an innate immune response.
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Progression of endothelial dysfunction in heart in a mouse model of atherosclerosis.
Filipová, Petra ; Němečková, Ivana (advisor) ; Doleželová, Eva (referee)
Charles University in Prague, Faculty od Pharmacy in Hradec Králové Department of Biological and Medical Sciences Progression of endothelial dysfunction in heart in a mouse model of atherosclerosis Author of diploma thesis: Petra Filipová Supervisor: RNDr. Ivana Němečková, Ph.D. Backround: Atherosclerosis is a disease when atherosclerotic lesions are formed in vessels. It represents the major cause of the heart failure and stroke. The endothelial dysfunction, which has been recorded in a large number of animals, is considered one of the main causes of atherosclerosis. In our work we have focused on the detection of the endothelial dysfunction markers in the heart of the ApoE/LDLR double knock-out mouse model. Methods: We detected changes in the expression of eNOS, p-eNOS and HO-1 in two-months and eight-months old ApoE/LDLR double knock-out mice, which were fed standard chow diet. For the semiquantitative evaluation of the observed markers we used the Western blot analysis of the homogenated left ventricle. Results: According to our results there is no difference in the expression of eNOS, p-eNOS and HO-1 in the studied mouse model of atherosclerosis in relation to age. This might mean, that the endothelial dysfunction in the coronary vessels of these mice doesn't develop. However, for further...
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