The aim of this work was demonstrate positive effect of n-3 polyunsaturated fatty acids EPA and DHA on the concentration of plasma triglycerides and cholesterol. According to the hypothesis, these polyunsaturated fatty acids are capable of reducing plasma cho-lesterol in mechanism, where is increased gene expression Insig-1 and simultaneously reduces the expression of genes encoding an enzyme for cholesterol synthesis (3-hydroxy-3-methylglutaryl-CoA reductase, a gene for HMG-CoA-R) and the gene for the LDL receptor, which is responsible for the entry of cholesterol from the plasma into the cells (low density lipoprotein receptor, the density, the LDL-R gene). LDL-R.The experiment was conducted on a model animal (Rattus norvegicus) fed with a standard feed mixture with the addition of 3% of salmon oil, fish oil 6% and 6% of oil from algae of the genus Schizochytrium. The control group was fed a feed mixture with the addition of 6% safflower oil, food with the addition of 3% palm oil was used as a negative control. Each group consisted of 10 animals. From the blood samples was determined levels of total cholesterol, HDL-cholesterol, LDL-cholesterol and triglycerides. The fatty acid content was determined in muscle, liver and adipose tissue. Food enriched with eicosapentaenoic acid and docosahexaenoic acid led to a reduction in total cholesterol and LDL-cholesterol levels. It was shown (P <0.05) after the addition of fish oil, salmon oil and algae of the genus Schizochytrium oil to feed mixtures of experimental rats, a decrease in total cholesterol (27%, 21%, 27% respectively) relative to controls. Our attempt partially confirmed the hypothesis. At the same time, we found that plasma lipid metabolism affects different mechanism than we thought.

The aim of this thesis was to observe how polyunsaturated fatty acids affect the wound healing process. This effect was observed by expression of selected genes and collagen type I and III in the model organism. The Wistar Albino laboratory rat strain was chosen as a model organism. 50 rats were used for the experiment. Rats were divi-ded into five groups of ten. One control group was selected which was fed only with compound feed without the addition of oil. The rats in the remaining groups were fed a feed mixture with the addition of oil (Schizochytrium extract oil, fish oil, palm oil and safflower oil). After 52 days of fattening, rats under anesthesia were excised on the dor-sal side of the back. Fattening was continued for 12 days and then rats were sacrificed with isoflurane to remove liver and healing skin wounds. EPA / DHA deposition was determined from collected liver. RNA was obtained from healing skin wounds for quan-titative PCR with specific primers for TGF-β1, PTGS2, ACTA2, VEGF, COL1A1, COL3A1. Further, immunohistochemical sections with type I and III collagen monoclo-nal antibodies were generated from the healing skin wound. The assumption of achie-ving better results with high DHA oil was not lost. Best results in the wound healing process were achieved in rats fed with safflower and fish oil (p <0.05). These results are inconsistent with the literature, since it was assumed that only n-3 PUFAs contribute to a better wound healing process and n-6 PUFAs, on the other hand, prolong wound hea-ling. Ambiguous results regarding the use of n-3 PUFAs in the wound healing process require further research.

Obesity, one of the most serious health problems of the 21st century, often occurs as a result of an imbalance between energy intake and energy expenditure. Dietary lipids play an important role in the development of obesity, partly because they represent the richest source of energy amongst all macronutrients. It is, however, not only the amount of consumed lipids, but also the composition of fatty acids, which strongly influences health effects of a particular diet. Saturated fatty acids (SFA) are generally considered as unhealthy due to their pro-inflammatory and lipotoxic properties, while monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA) represent a healthier alternative, as they are more readily oxidized and do not disrupt biochemical properties of cellular membranes. Amongst PUFA, PUFA of n-3 series (Omega-3) represent an utterly unique class of lipids that have been documented to protect against cardiovascular disease and dyslipidemia in men and improve insulin sensitivity and glucose tolerance primarily in animal models of obesity. Some molecular mechanisms of Omega-3 action have been already uncovered, such as the modification of biological membranes composition, activation of various transcription factors and membrane receptors, and their role as precursors for...

Obesity and associated metabolic disorders, called as "metabolic syndrome", currently represent a major social and economical problem of public health. From the energy balance point of view, long-lasting energy surplus leads eventually to massive accumulation of energy stores resulting in various adverse effects on metabolism and health. General goal of the thesis was to examine these metabolic disorders at cellular and whole-body level using suitable mouse models. The main focus was on the most metabolically active tissue, namely skeletal muscle, liver and adipose tissue and on the regulatory roles of AMP-activated protein kinase (AMPK) and leptin in the energy metabolism. The whole thesis is based on four published studies. Two studies were focused on skeletal muscle. In the first study, we proved the involvement of leptin and AMPK in the metabolic response to high-fat diet-feeding. We described a mechanism of muscle non- shivering thermogenesis based on enhanced lipid catabolism, which contributes to the genetically-determined resistance of inbred A/J mice to obesity. Such mechanism was not operating in obesity-prone C57BL/6 mice. In the second study, performed using C57BL/6 mice, we have described beneficial effect of combination treatment using n-3 polyunsaturated fatty acids (n-3 PUFA) of...

The important factor in regulation of metabolic processes is regulatory proteins, which are able to react by feed-back to energetic state of the cell. Big attention is focused on the AMP activated kinase (AMPK) and NAD+ activated deacetylase SIRT1. These enzymes interact together and their stimulation increases mitochondrial biogenesis and fatty acid oxidation. Due to this it functions beneficially against the onset of obesity, insulin resistance and ageing. Fasting, exercise and some antidiabetogenic drugs act by these regulators. n-3 polyunsaturated fatty acids (PUFA) are also known because of their stimulative effects on mitochondrial biogenesis and -oxidation. Previous work of our group have showed that intake of higher dose of n-3 polyunsaturated fatty acids (PUFA) in diet lead to increase in activity of AMPK in white adipose tissue. New results presented in this thesis show that SIRT1 is essential for increase in expression of stimulators of -oxidation (PPAR etc) in response to n-3 PUFA in diet. n-3 PUFA futher improve the metabolic profile synergistically with calorie restriction probably through SIRT1.

Charles University Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Candidate: Monika Novotná Supervisor of Diploma Thesis: Mgr. Monika Kuchařová, Ph.D. Title of Diploma Thesis: The determination of fatty acid levels in the tissues of healthy, suddenly deceased persons and polymorbid patients The aim of the thesis was to determine the levels of 14 fatty acids in tissues of suddenly deceased, otherwise healthy individuals and in the group of polymorbid, chronic patients. It was a comparison of fatty acid levels in seven tissues of the human body: subendocardial left ventricular tissue, liver parenchyma tissue, kidney cortex, adrenal tissue, skeletal muscle, abdominal subcutaneous adipose tissue, and brain tissue. Each group included 10 deceased patients. The theoretical part incudes fatty acids as the main component of lipids. It deals mainly with the group of polyunsaturated fatty acids and their relation to pathologies in the human body. Gas chromatography, thanks to which we analyzed the fatty acid spectrum, is also described. The experimental part consists of the basic characteristics of the research groups, the description of the workflow and the results. Statistically processed data are divided into categories by individual fatty acids and other monitored...

Obesity is tightly connected with metabolic diseases including insulin resistance, type 2 diabetes or dyslipidemia. Peroxisome proliferator-activated receptor (PPAR)-α is a key transcription factor involved in the regulation of lipid metabolism, while its activity is stimulated by a variety of hypolipidemic drugs. n-3 polyunsaturated fatty acids (PUFA), including eicosapentaenoic (EPA) and docosahexaenoic (DHA) acid, are endogenous ligands of PPARα, and they are used in the form of fish oil as dietary supplements in order to lower blood lipid levels and to prevent cardiovascular disease. Wax esters represent a novel lipid form of EPA and DHA, and according to recent studies they could exert more potent effects than the classical fish oil (i.e. triacylglycerols). Mice of the 129S1/SvImJ inbred strain were used in the present experiment, and included wild-type (WT) mice, as well as transgenic mice either with the exclusive expression of the human form of PPARα (hPPARα) or mice completely lacking PPARα (PPARα-KO). Mice were fed for 8 weeks the following diets: (i) a control low-fat diet, (ii) obesogenic high-fat diet (cHF), and (iii) the cHF diet supplemented with the n-3 PUFA concentrate in the form of wax esters isolated from marine zooplankton Calanus finmarchicus (ω3Cal). Mice were subjected to...

Regulation of the flow of fatty acids (FA) by futile cycle (TAG/FA cycle) in white adipose tissue (WAT) is an important mechanism of controlling metabolism of FA and therefore its regulation is in the interest of research as a possible therapeutic target in the treatment of obesity and insulin resistance. The study of the murine model suggests that the treatment of the n-3 PUFA with mild caloric restriction (CR) probably induces the TAG/FA cycle mainly in epididymal depot. It also reduces inflammation in WAT and the risk of cardiovascular diseases. We focused on monitoring the gene expression encoding key enzymes of the TAG/FA cycle (PEPCK, ATGL, HSL, DGAT1 and DGAT2) in dorzolumbar (DL), gonadal (GON) and mesenteric (MEZ) depot in a murine model C57BL/6 using qPCR. We were interested in the influence of the composition of fats in the diet, the influence of administration of n-3 PUFA with combination of 10% CR and the difference of gene expression among depots of WAT. The results indicate that the supplementation of high-fat diet with n-3 PUFA contributes mainly to reduction of gene expression for DGAT2 and the combination of n-3 PUFA and CR increases expression of genes influencing the TAG/FA cycle. In MEZ unlike GON and DL fat depot were no significant differences in gene expression, while the different...

Charles University Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Author of Diploma Thesis: Bc. Barbora Kuříková Supervisor of Diploma Thesis: Mgr. Monika Kuchařová, Ph.D. Consultant: prof. MUDr. Zdeněk Zadák, CSc. Title of Diploma Thesis: Determination of the fatty acids spectrum in patients undergoing treatment with taxanes The diploma thesis deals with the determination of fatty acids spectrum in erythrocytes in patients treated with taxanes. The theoretical part describes general features of fatty acids, their synthesis and degradation. Then it is focused on the topic of breast cancer, taxane treatment and negative side effects associated with taxane treatment, especially polyneuropathy. The gas chromatography, which is commonly used in practice, is also described in this part. The experimental part is divided into three parts. The first part describes working process, chromatographic analysis and evaluation of this analysis. In the second part there is comparing of spectrum of fatty acids of patients without and with polyneroupathy. And in the last part there is a comparing of spectrum of fatty acids of patients before taxane treatment, closely after treatment and about month after the treatment. Results of the experimental part are evaluated at the end...

Charles University Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Candidate: Alena Čunátová Supervisor of Diploma Thesis: Mgr. Monika Kuchařová, Ph.D. Title of Diploma Thesis: The fluctuation of fatty acids levels in the tissues of suddenly deceased persons and comparison with values in pathological states Fatty acids and their metabolites are significantly involved in many physiological and pathological processes. This thesis monitors the levels of selected fatty acids in human tissues and the effect of long-term disease on their stores. In addition to the general characteristics, the theoretical part focuses on the intake of polyunsaturated fatty acids and their importance in the human body. It also deals with the effects of eicosanoids and other fatty acid metabolites. Methods used in fatty acid bioanalysis are also described. In the experimental part, the proportion of fatty acids in seven tissue types was compared between two groups of donors. One of them included 8 relatively young, healthy, suddenly deceased individuals. The second group included 12 elderly polymorbid patients. The obtained tissue samples were adjusted using extraction and derivatization techniques. Gas chromatography with flame ionization detection was used for analysis. The measured...