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Chemical and biochemical transformation of bioactive compounds
Šimášková, Ema ; Sokolová, Romana (advisor) ; Martínková, Markéta (referee)
Xenobiotics, such as pharmaceuticals, food additives, environmental pollutants, and dietary bioactive compounds in organism are metabolized by various enzymes, resulting in their bioactivation or detoxification. Identification of structure of resulting metabolites is important for their detection in bodily fluids and tissues for diagnostic and forensic purposes. This thesis reviews known biochemical processes and enzymes involved in xenobiotic metabolism, including cytochromes 450 (CYP) and flavine monooxygenases (FMO). Given that biochemical reactions are to a major extent composed of electron-transfer reactions (i.e. oxidation and reduction), the thesis includes a section dealing with the practical approaches to determination of the oxidative or reductive mechanism of bioactive compounds.
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Development of novel Constitutive androstane receptor (CAR) ligands
Dušek, Jan ; Pávek, Petr (advisor) ; Hudeček, Jiří (referee) ; Chládek, Jaroslav (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate Mgr. Jan Dušek Supervisor Prof. PharmDr. Petr Pávek, Ph.D. Title of Doctoral Thesis Development of novel Constitutive androstane receptor (CAR) ligands Constitutive androstane receptor is nowadays known as the established nuclear receptor that regulates the expression of several key cytochrome P450 enzymes, predominantly CYP3A4 and CYP2B6. Recently it has been shown that CAR has also essential role in the regulation of endogenous metabolism of glucose, lipids, cholesterole or bile acids. Simultaneously, this receptor is considered to have proliferative effect on human hepatocytes and protective effects against toxic and dietary damage of liver parenchyme. Given the possible therapeutic utilization of CAR, its therapeutic options are being intensively studied. Unfortunately, currently known ligands of human or mouse CAR are either poorly selective or indirect. The aim of my doctoral thesis was to find new ligands of mouse and human CAR, that would enable more detailed study of the receptor.
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Effect of dihydromyricetin on ethanol metabolism
Skotnicová, Aneta ; Hodek, Petr (advisor) ; Koblas, Tomáš (referee)
Dihydromyricetin (DHM), also ampelopsin, is a flavonoid compound which exhibits a broad spectrum of positive effects on the human body. Herbal extracts containing this compound have been widely used in traditional Chinese medicine mainly for their hepatoprotective properties. DHM also helps with alcohol intoxication and reduces the signs of hangover or abstinence. Given the fact that the mechanism of DHM effects on the ethanol metabolism has not been clarified yet, the effect of dihydromyricetin on the expression and activity of alcohol dehydrogenase (ADH), one of the most important enzymes involved in ethanol metabolism, was therefore studied in this thesis. The cultivation conditions of primary hepatocytes which were isolated from unpretreated and ethanol-pretreated rats and subsequently exposed to EtOH and DHM were optimized. While determining the degree of cell damage caused by EtOH in the presence of DHM, no significant trend in the protective effect of DHM was found. On the other hand, the protective effect of ethanol in hepatocytes cultivated in EtOH and DHM was detected by technique of ELISA (the determination of alanine transaminase). The Western blot technique followed by immunodetection did not detect the induction of ADH expression in hepatocytes. Furthemore, the modulation effect of...
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Effect of iron overload on the induction of apoptosis in mammalian cells
Kabíčková, Tereza ; Balušíková, Kamila (advisor) ; Klíma, Martin (referee)
Iron cations are an important metal ions required to number of essential cell functions. On the other hand, ferrous iron can be very toxic as well. When surplus iron is present in cells, it can catalyze the formation of reactive oxygen species (especially hydroxyl radicals) by Fenton reaction. Iron homeostasis is predominantly regulated by very strict mechanisms on the level of iron uptake into the body. Moreover, iron absorption, transport and storage within the body can be also regulated using complex mechanisms which differ on the level of individual cells and on the level of whole organism. Deregulation of iron homeostasis causing an iron overload and generation of reactive oxygen radicals can evoke serious cell damage leading up to apoptotic cell death. Excess iron storage and subsequent development of oxidative stress can affect lot of different tissues in the body. The organ damages such as fibrosis, cirrhosis, hepatocellular carcinoma, heart failure, loss of β cells and glucose intolerance or diabetes mellitus in patients with iron overload are very often seen. Nevertheless, the apoptosis induced by iron overload has not been well elucidated yet. There are no complex informations about the precise mechanism by which oxidative stress affects different cell types or whether there are other...
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Pharmacological modifications of potential signal systems regulating metabolism of adipocytes and hepatocytes and their influence on obesity
Hodis, Jiří ; Farghali, Hassan (advisor) ; Kršiak, Miloslav (referee) ; Otová, Berta (referee)
v anglickém jazyce: Thesis abstract: Background and aims: Both obesity and metabolic syndrome form severe health problems in the whole world. Nevertheless the armament of pharmacotherapy for both diseases remains unsatisfactory. We aimed our work to main organs in risk of the mentioned diseases -liver and visceral fat using hepatocytes and visceral adipocytes as model. We detected 3 main metabolic and signalization activities- glycogenolysis, Nitric oxide (NO) production and transcription of inducible NO synthase (iNOS) in hepatocytes, lipolysis, NO production and iNOS transcription rate in adipocytes. We directed our interest to combination of peroxisome proliferation activator receptor γ (PPARγ) agonist, antagonist and β3 adrenergic agonist in the culture of epididymal rat adipocytes in the first part of our work. While in the second part we investigated the influence of β and α adrenergic mimetics, adrenergic blockers in the culture of rat high glycogen content hepatocytes. Methods: NO production was detected under the active agents treatments by detection of NO oxidative products NO2 and NO3 in media. Glycogenolysis was measured as free glucose rise released by hepatocytes into the media. NOS transcription level was extrapolated after comparative polymerase chain reaction with reverse...
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